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1.
AIM: To investigate the role of imperatorin in reversing the resistance of the PC9 CD133+ cell subsets to gefitinib. METHODS: MTT assay was performed to evaluate the viability of PC9 cells treated with imperatorin and gefitinib. The expression of c-met, activation of caspases and phosphorylation of epidermal growth factor receptor (EGFR), PI3K and AKT in the PC9 cells treated with imperatorin and gefitinib were determined by Western blot. The percentage of CD133+ cell subsets population and the apoptotic rate of the PC9 cells treated with imperatorin and gefitinib were analyzed by flow cytometry. RESULTS: The sensitivity of the PC9 CD133+ cell subsets to gefitinib was significantly lower than that of the PC9 CD133- cell subsets. Treatment with gefitinib alone significantly inhibited the protein levels of EGFR/PI3K/AKT in the PC9 CD133- cell subsets but not the PC9 CD133+ cell subsets. Treatment with gefitinib alone increased the percentage of CD133+ cell subsets population in the PC9 cells. However, combination of gefitinib with imperatorin significantly inhibited the enrichment of CD133+ cell subsets population. Imperatorin down-regulated c-met expression, suggesting the c-met was the target of imperatorin in the PC9 CD133+ cell subsets. The results of MTT assay, Western blot analysis and flow cytometry indicated that imperatorin increased the gefitinib induced inhibition of PI3K/AKT protein levels by down-regulating the expression of c-met, which subsequently induced the cleavage of caspases and apoptosis in the PC9 CD133+ cell subsets.CONCLUSION: Imperatorin increases the sensitivity of lung cancer CD133+ cell subsets to gefitinib by down-regulating the expression of c-met, and the synergistic anti-tumor effect exists between imperatorin and gefitinib. 相似文献
2.
AIM:To investigate the effect of metformin combined with paclitaxel on the viability and apoptosis of breast cancer cell line MCF-7 and its possible mechanism. METHODS:MCF-7 cells were treated with metformin at different concentrations (2, 5, 10, 20, 40 and 80 mmol/L) and in vitro cultured. The viability of MCF-7 cells was measured by MTT assay. Metformin at 2 mmol/L or paclitaxel at 2.4 mg/L alone or in combination was used to treat the cells, and compound C, an inhibitor of adenosine monophosphate-activated protein kinase (AMPK) signaling transduction pathway, was also used. The cells were divided into control group, metformin group, paclitaxel group, combination group, and combination +compound C group. The apoptosis of the cells was analyzed by flow cytometry. The expression of Bax, Bcl-2 and caspase-3 at mRNA and protein levels was determined by RT-qPCR and Western blot. The protein levels of AMPK and P21 were examined by Western blot. RESULTS:Metformin at different concentrations (2, 5, 10, 20, 40 and 80 mmol/L) significantly inhibited the cell viability in a concentration-dependent manner (P<0.05). Compared with control group, treatment with metformin at 2 mmol/L or paclitaxel at 2.4 mg/L alone or in combination significantly inhibited the cell viability, induced apoptosis (P<0.05), decreased the level of Bcl-2 (P<0.05), increased the levels of Bax and caspase-3 (P<0.05), and promoted the protein expression of AMPK and P21 (P<0.05). The effects of metformin and paclitaxel in combination were better than those of single drug treatment, while AMPK inhibitor weaken these effects. CONCLUSION:Metformin combined with paclitaxel inhibits the viability and induces the apoptosis of breast cancer MCF-7 cells by activating AMPK signaling pathway and regulating apoptosis signaling pathway. 相似文献
3.
AIM: To study the effect of histone deacetylase 1 (HDAC1) on the apoptosis of breast cancer cells.METHODS: The expression of HDAC1 at mRNA and protein levels in normal mammary epithelial cell line MCF-10A and breast cancer cell lines BT549, MCF-7 and MDA-MB-231 was measured by RT-qPCR and Western blot. HDAC1 siRNA was transfected into MDA-MB-231 cells, and then RT-qPCR and Western blot were used to determine the expression level of HDAC1. The cell viability was measured by MTT assay, and apoptosis was analyzed by flow cytometry. The protein levels of β-catenin, c-Myc, cyclin D1 and cleaved caspase-3 were determined by Western blot. Breast cancer cells with HDAC1 knockdown were treated with Wnt/β-catenin signaling pathway activator, and then the cell viability and apoptosis were measured.RESULTS: The expression of HDAC1 at mRNA and protein levels in BT549, MCF-7 and MDA-MB-231 cells was significantly higher than that in normal mammary epithelial cell line MCF-10A, and the highest expression level of HDAC1 was observed in MDA-MB-231 cells (P<0.05). HDAC1 siRNA reduced the expression of HDAC1 at mRNA and protein levels in the breast cancer cells. The viability of MDA-MB-231 cells was decreased after knockdown of HDAC1 expression, the apoptotic rate was increased, the protein level of cleaved caspase-3 in the cells was elevated, and the protein levels of β-catenin, c-Myc and cyclin D1 were decreased (P<0.05). Wnt/β-catenin signaling pathway activator reversed HDAC1 knockdown-induced apoptosis and decrease in viability of MDA-MB-231 cells, and reduced the protein level of cleaved caspase-3.CONCLUSION: Knockdown of HDAC1 expression induces apoptosis of breast cancer cells by inhibiting the activation of Wnt/β-catenin signaling pathway. 相似文献
4.
AIM: To construct a prokaryotic expression plasmid to produce recombinant human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and to verify the biological activity of TRAIL. METHODS: The prokaryotic expression plasmid pET-28a (+)-TRAIL114-281 was constructed. Human soluble TRAIL was obtained through optimized inducing protein expression and purification conditions. The biological activity of TRAIL was verified by CCK-8 assay. The apoptosis-inducing effect of TRAIL alone and/or in combination with proteasome inhibitor bortezomib (Velcade, PS-341) on the tumor cell lines H460(TRAIL-sensitive) and K562(TRAIL-resistance) for 24 h was determined. The apoptotic rates of the cells were analyzed by flow cytometry with Annexin V-FITC/PI staining. The activities of caspase-8, -9 and -3 in the cells were detected by colorimetric method. The protein expression of Bax, Bcl-2 and cFLIP was measured by Western blot. The expression of DR4 and DR5 in the H460 cells and K562 cells after treated with bortezomib for 24 h was detected by flow cytometry. RESULTS: The recombinant human soluble TRAIL protein with stable bioactivity was successfully acquired, which induced apoptosis in H460 cells and K562 cells. After treatment with different concentrations of TRAIL, the apoptotic rate of H460 cells was significantly increased with the increase in the concentration of TRAIL (P<0.05), but the apoptotic rate of K562 cells was not affected by the increasing TRAIL concentration. Apoptotic rate in combination group was obviously higher than that in single group (P<0.05). In the process of apoptosis, the activities of caspase-8, -9 and -3 in H460 cells and K562 cells were both increased. The expression of Bcl-2 and cFLIP in treatment groups (especially the combination group) was decreased compared with control group. No significant change of the Bax expression level was observed. The expression of DR4 and DR5 in the H460 cells and K562 cells was significantly up-regulated after treated with bortezomib (P<0.05). CONCLUSION: Bortezomib combined with recombinant human soluble TRAIL synergistically induces apoptosis in tumor cell lines H460 and K562 through initiating intrinsic apoptotic pathways by up-regulating death receptors DR4 and DR5, and reducing the expression of antiapoptotic proteins Bcl-2 and cFLIP. 相似文献
5.
ZHANG Qing-ran ZHOU Zhao-ling PAN Zhen-hai MA Ya-peng MA Zhi-qiang FEI Hong-rong 《园艺学报》2018,34(2):212-217
AIM: To investigate the effects of evodiamine on the growth and apoptosis of human hepatocellular carcinoma Huh7 cells, and to illustrate the molecular mechanism that evodiamine enhances antitumor activity of tumors necrosis factor-related apoptosis-inducing ligand (TRAIL) in Huh7 cells.METHODS: The cell viability was measured by MTT assay. The cell cycle distribution was analyzed by flow cytometry. The apoptosis rate was determined by TUNEL staining. The protein levels of cell cycle-and apoptosis-related proteins were detected by Western blot analysis.RESULTS: Treatment of Huh7 cells with evodiamine reduced the cell viability (P<0.05). Evodiamine induced cell cycle arrest in G2/M phase by upregulation of p27, cyclin B1, cell division cycle protein 2 (Cdc2) and p-Cdc2. Evodiamine triggered apoptosis accompanied by cleavage of caspase-3 and poly(ADP-ribose) polymerase (PARP). Combination of evodiamine with TRAIL significantly reduced the cell viability and increased cleavage of caspase-3 and PARP as compared with the use of each agent alone. Moreover, evodiamine increased the expression of death receptor 5 (DR5) in the Huh7 cells.CONCLUSION: Evodiamine inhibits the cell growth by reducing the cell viability and inducing cell cycle arrest. Evodiamine also triggers cell apoptosis and enhances the sensitivity of Huh7 cells to TRAIL by upregulating the expression of DR5. 相似文献
6.
AIM: To detect the expression of WNT5B in normal breast epithelial cells and different breast cancer cell lines, and to investigate the effects of WNT5B over-expression on the viability and apoptosis of human breast cell line MCF-7.METHODS: The mRNA expression of WNT5B was detected by RT-PCR in different breast cancer cells. MCF-7 cells were transfected with plasmid pcDNA3.1/WNT5B or pcDNA3.1, and the expression of WNT5B at mRNA and protein levels was examined in the 2 groups by real-time PCR and Western blotting, respectively. Subsequently, the changes of cell viability and cell apoptosis were analyzed by CCK-8 assay and flow cytometry, respectively. RESULTS: The expression of WNT5B in the breast cancer cell lines was lower than that in MCF10A cells. The WNT5B expression in the MCF-7 cells in experimental group was significantly higher than that in vector group (P<0.05). However, the cell viability in experimental group decreased significantly as compared with vector group (P<0.05). The number of the cells in S-phase obviously increased, while the percentage of the cells in G1-phase and G2/M-phase decreased compared with vector group. The number of apoptotic cells in WNT5B group was significantly higher than that in vector group.CONCLUSION: The expression of WNT5B is decreased in breast cancer cells. WNT5B over-expression significantly inhibits the cell growth and promotes the cell apoptosis in breast cancer MCF7 cells. 相似文献
7.
AIM: To study the effect of poly(ADP-ribose) polymerase-1 (PARP-1) on cisplatin resistance of human breast cancer MCF-7 cells and its possible mechanisms.METHODS: The expression of PARP-1 at mRNA and protein levels in MCF-7 cells and MCF-7/DDP cells was determined by real-time PCR and Western blot. The expression of PARP-1 in the MCF-7/DDP cells was blocked by PARP-1 siRNA. The cell viability and apoptosis were detected by the CCK-8 assay and flow cytometry analysis, respectively. Furthermore, the protein levels of PARP-1, Bcl-2, Bax, cleaved caspase-3, caspase-3, cytochrome C (Cyto-C), extracellular signal-regulated kinase (ERK) and phosphorylated ERK (p-ERK) were detected by Western blot.RESULTS: The expression of PARP-1 at both mRNA and protein levels was significantly up-regulated in the MCF-7/DDP cells. The expression of PARP-1 was increased in the MCF-7 cells treated with cisplatin. Knockdown of PARP-1 induced the apoptosis of MCF-7/DDP cells with an increased sensitivity to cisplatin. Meanwhile, knockdown of PARP-1 down-regulated the protein levels of Bcl-2/Bax and p-ERK, but up-regulated the protein levels of cleaved caspase-3 and Cyto-C. After incubated with a specific ERK inhibitor U0126, the cell viability in PARP-1 siRNA group was down-regulated significantly.CONCLUSION: Knockdown of PARP-1 increases the sensitivity of MCF-7/DDP cells to cisplatin, and promotes the cell apoptosis via mitochondrial apoptosis pathway. The mechanism may be related to the attenuation of ERK signaling pathway by inhibiting phosphorylation of ERK. 相似文献
8.
XU Tong-peng LIU Li-gen JIANG Xue-wei GAO Wu XIE Ying-hua ZHAO Li-min HAN Xi-yao LU Dao-pei 《园艺学报》2011,27(3):481-487
AIM: To investigate the effect of plumbagin and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the apoptosis of leukemic Kasumi-1 cells. METHODS: Kasumi-1 cells were treated with plumbagin alone, recombinant soluble TRAIL(rsTRAIL) alone or the combination of plumbagin with rsTRAIL to induce apoptosis. The cell proliferation was analyzed by CCK-8 assay. Apoptosis was determined by flow cytometry with AnnexinⅤ/PI double staining and TUNEL staining. The expression of DR4 and DR5 at mRNA level was measured by real-time PCR. The expression of signal transduction proteins, such as DR5, caspase-3, caspase-8, caspasep-9, Bid, Bax and c-FLIP was detected by Western blotting. RESULTS: Both rsTRAIL and plumbagin induced the apoptosis in Kasumi-1 cells, and combination of plumbagin with rsTRAIL enhanced the apoptosis. The ratios of Annexin V-positive Kasumi-1 cells were (27.7±2.9)%, (25.6±3.1)% and (52.1±3.3)% in 100 μg/L rsTRAIL group, 2 μmol/L plumbagin group and the combination group, respectively, and the positive rate in combination group was significantly higher than those in other 2 groups. TUNEL assay demonstrated that the number of apoptotic cells in combination group was higher than that in the cells treated with rsTRAIL or plumbagin alone. Plumbagin up-regulated the expression of DR5 at mRNA level in Kasumi-1 cells, and up-regulation of DR5, activation of caspase-8 and down-regulation of c-FLIP at protein level were detected in the cells treated with plumbagin alone and the combination of plumbagin with rsTRAIL. CONCLUSION: Plumbagin enhances TRAIL-induced apoptosis in Kasumi-1 cells by up-regulating DR5, activating caspase-8 and down-regulating c-FLIP. 相似文献
9.
AIM: To investigate the apoptotic pathway of MCF-7 breast cancer induced by the grub extract in vitro.METHODS: MTT assay was used to determine the effect of the grub extract on proliferation of MCF-7 human breast cancer cell line and cell toxicity. Morphological changes of the apoptosis in cancer cells were observed by HE staining through invert microscope, light microscope, AO/EB double fluorescent staining under fluorescent microscope. FCM was used to assay the change of apoptotic rate. The expression of Bcl-2, Fas, caspase-9, caspase-3 in apoptotic pathway was detected with immunocytochemical method before and after exposure to the grub extract, and the effect of that on apoptotic pathway was explored.RESULTS: (1) The MTT test showed that the growth of MCF-7 human breast cancer cell line was significantly inhibited by the grub extract in dose and time dependent manners. The inhibitory rate in exposure group was significantly different from that in control group (P<0.01). (2) Morphological changes of apoptosis including nuclear condensation, fragment and apoptosis body formation were observed by invert microscope. (3) The MCF-7 human breast cancer cells in experimental group by HE staining showed nuclear condensation and blue-black, cytoplasm slight red, nuclear chromatin condensation and fragment shape, apoptosis body formations. (4) Apoptosis in the experimental group was observed by AO/EB double fluorescent staining under fluorescent microscope. (5) FCM assay indicated that apoptotic rate increased significantly in time dependent manner in experimental group. (6) The expression of Bcl-2 was down-regulated, while that of Fas, caspase-3, caspase-9 was up-regulated, compared with control group (P<0.01).CONCLUSION: (1) The proliferation of MCF-7 human breast cancer cell line can be inhibited significantly by the grub extract in vitro. (2)The mechanism of effect of the grub extract on MCF-7 human breast cancer cell line might be mediated by down-regulation of Bcl-2 and up-regulation of Fas, caspase-3, caspase-9. This type of apoptosis starting and performing is through death receptor pathway and mitochondrial pathway. 相似文献
10.
AIM: To investigate the inhibitory effect of CC-223, an inhibitor of mammalian target of rapamycin (mTOR) kinase, on the viability of human breast cancer cells and its mechanism. METHODS: The inhibitory effect of CC-223 on the viability of MCF-7 cells and MDA-MB-231 cells was measured by CCK-8 assay. The cell cycle distribution of breast cancer cells was examined by flow cytometry. The expression of cell cycle-related proteins and oncoproteins c-Myc and survivin was analyzed by Western blot. RESULTS: CC-223 significantly inhibited the viability of both MCF-7 and MDA-MB-231 cells (P <0.05). CC-223 induced cell cycle arrest in both G1 phase and G2/M phase in the MCF-7 cells (P <0.05). However, low concentration of CC-223 treatment resulted in the accumulation of MDA-MB-231 cell cycle in the G2/M phase, and the cell number in G1 phase was unaffected. Treatment with CC-223 for 24 h clearly inhibited the protein levels of cyclin B1, cyclin D1 and phosphorylated cell division cycle protein 2 in the breast cancer cells (P <0.05). CC-223 suppressed the expression of c-Myc and survivin in both MCF-7 cells and MDA-MB-231 cells (P <0.05). CONCLUSION: CC-223 inhibits cell viability by blocking cell cycle progression and down-regulating expression of c-Myc and survivin in both MCF-7 and MDA-MB-231 cells. 相似文献
11.
12.
AIM: To investigate the effect of Notch1 gene silencing on phosphorylations of JNK1 and p53 in human breast cancer MCF-7 cells.METHODS: shRNA-Notch1 eukaryotic expression plasmid was constructed and transfected into MCF-7 cells. The expression of Notch1 and Hes-1 was observed by Western blotting after transfction. Apoptosis and mitochondrial membrane potential were detected by flow cytometry. Western blotting was also used to determine the protein levels of p-JNK1, p-p53, PUMA, NOXA and cleaved caspase-3 after Notch1 silencing was performed in MCF-7 cells.RESULTS: Silencing of Notch1 significantly reduced the expression of Notch1 and Hes-1 in MCF-7 cells (P<0.01). In shNotch1 group, the number of apoptotic cells was much higher (P<0.01) and mitochondrial membrane potential was much lower (P<0.05) than those in shControl group. The protein levels of p-JNK1, p-p53, PUMA, NOXA and cleaved caspase-3 increased obviously after silencing of Notch1 was performed in MCF-7 cells (P<0.05).CONCLUSION: Notch1 silencing induces apoptosis of human breast cancer MCF-7 cells through promoting phosphorylations of JNK1 and p53, and increasing the production of PUMA, NOXA and cleaved caspase-3. 相似文献
13.
AIM:To study the effects of sodium aescinate on the apoptosis of cervical cancer HeLa cells and its molecular mechanism. METHODS:MTT assay was used to detect the growth and proliferation of HeLa cells. The morphological alteration was observed under inverted microscope. Annexin V-FITC/PI double staining and DAPI nuclear staining were used to determine the apoptosis of HeLa cells induced by sodium aescinate. The apoptosis-related proteins PARP, cleaved caspase-8 and pro-caspase-3, and the proliferation-associated molecules Akt and ERK, as well as TRAIL receptors DR4 and DR5 were detected by Western blotting. RESULTS:Sodium aescinate inhibited the growth of HeLa cells in a concentration-dependent manner. Treatment with sodium aescinate induced the typical morphology of apoptotic cells and increased the apoptotic rate significantly. The cleaved PARP, cleaved caspase-8 and cleaved caspase-9 protein expression was observed. The expression of DR4 and DR5 was up-regulated. Meanwhile, pro-caspase-3 was decreased, and the levels of p-Akt and p-ERK were down-regulated by sodium aescinate in a dose-dependent manner. CONCLUSION:Sodium aescinate inhibits the proliferation and promotes the apoptosis of HeLa cells by increasing death receptor expression and repressing proliferation-associated signaling pathways. 相似文献
14.
Artesunate enhances anti-tumor activity of gemcitabine on pancreatic cancer through MDM2/p53 pathway
AIM: To explore the effect of artesunate on gemcitabine-dependent anti-tumor activity for pancrea-tic cancer.METHODS: The viability of p53 wild-type pancreatic cancer cell line Capan-2 was measured by MTT assay. The protein levels of murine double minute protein 2 (MDM2), p53, Noxa and Puma, the release of cytochrome C and apoptosis-inducing factor, and the activation of caspase-9 and caspase-3 in the Capan-2 cells were determined by Western blot. The mitochondrial membrane potential and apoptotic rate of the Capan-2 cells were analyzed by flow cytometry.RESULTS: The viability of Capan-2 cells in artesunate plus gemcitabine group was significantly lower than that in single gemcitabine treatment group (P<0.05). Gemcitabine treatment significantly inhibited the expression of MDM2 in the Capan-2 cells. The expression levels of p53, Noxa and Puma in the artesunate plus gemcitabine group were significantly higher than those in single gemcitabine treatment group. Artesunate obviously promoted the apoptosis, the release of cytochrome C and apoptosis-inducing factor, the activation of caspase-9 and caspase-3, and the collapse of mitochondrial membrane potential in the Capan-2 cells treated with gemcitabine. On the other hand, transfection with MDM2 plasmid obviously suppressed the apoptotic pathway of Capan-2 cells which were treated with artesunate and gemcitabine.CONCLUSION: Artesunate enhances the anti-tumor activity of gemcitabine on pancreatic cancer Capan-2 cells through the MDM2/p53 pathway. 相似文献
15.
AIM:To investigate the autophagy of breast cancer cells induced by baicalein and to explore its mechanism.METHODS:The effects of baicalein on the viability of MCF-7 cells and 4T1 cells were investigated by MTT assay,and the dosage of the drug was determined.The expression levels of microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ) and LC3-I in the MCF-7 cells and 4T1 cells treated with baicalein at doses of 25,50 and 100 μmol/L,or combined with autophagy inhibitor 3-methyladenine (3-MA) were determined by Western blot.In order to confirm the role of baicalein in autophagy,the effect of 3-MA on the apoptosis of both MCF-7 cells and 4T1 cells induced by baicalein was analyzed by flow cytometry.The protein levels of p-mTOR,mTOR,p-AKT and AKT were examined by Western blot and the role of AKT-mTOR pathway in the induction of autophagy in breast cancer induced by baicalein was determined by the combination of activators.RESULTS:Baicalein at 50 μmol/L and above doses significantly inhibited the viability of breast cancer cells in a dose-and time-dependent manner.The expression of LC3-Ⅱ/LC3-I in both MCF-7 cells and 4T1 cells was significantly enhanced after the action of baicalein,and the ratio of LC3-Ⅱ/LC3-I was significantly decreased after 3-MA addition.The results of flow cytometry showed that,compared with baicalein group,the combination of baicalein and 3-MA promoted the levels of necrosis and apoptosis.Moreover,the protein levels of p-mTOR and p-AKT were significantly decreased and were rescued by EGF,while their total protein levels were not changed.CONCLUSION:Baicalein induces autophagy through AKT-mTOR pathway both in MCF-7 cells and 4T1 cells. 相似文献
16.
AIM: To determine whether imperatorin would enhance the effect of doxorubicin therapy on cervical cancer in vitro.METHODS: The viability of HeLa cells treated with imperatorin and doxorubicin was determined by MTT assay in vitro. The expression of Bcl-2 protein family(Mcl-1, Bcl-2, Bcl-xL, Bad and Bax) in HeLa cells treated with imperatorin and doxorubicin was evaluated by Western blot analysis. The apoptosis and mitochondrial membrane potential(ΔΨm) in the HeLa cells treated with imperatorin and doxorubicin were analyzed by flow cytometry. A Mcl-1 expression vector was constructed, and its role in the cytotoxicity of imperatorin plus doxorubicin to HeLa cells was detected by MTT assay.RESULTS: Addition of imperatorin significantly enhanced the cytotoxicity of doxorubicin to HeLa cells in vitro. Mcl-1 expression was down-regulated by imperatorin but was not influenced by doxorubicin in the HeLa cells. A combination of imperatorin and doxorubicin induced apoptosis and ΔΨm collapse more significantly compared with the treatment with imperatorin or doxorubicin alone. Furthermore, the imperatorin-induced sensitization for doxorubicin cytotoxicity to HeLa cells was abolished by the transfection with Mcl-1 expression plasmid.CONCLUSION: The combination of doxorubicin with imperatorin enhances the antitumor effect of doxorubicin on cervical cancer cells via targeting Mcl-1. 相似文献
17.
AIM: To explore the effect of new artificially synthesized androgen receptor (AR) antagonist HC-1119 on the biological function of triple-negative breast cancer (TNBC) BT549 cells and the molecular mechanism. METHODS: The AR expression was assessed in different human breast cancer cell lines MDA-MB-231, T47D, MCF-7, SKBR3 and BT549 by Western blot. The TNBC BT549 cells with AR positive expression were treated with HC-1119. The cell viability was measured by CCK-8 assay. The apoptosis rate and cell cycle distribution were analyzed by flow cytometry. The migration and invasion abilities were detected by Transwell assay in vitro. The protein expression of E-cadherin, vimentin and P21 was determined by Western blot. RESULTS: AR was positively expressed in BT549 cells. HC-1119 inhibited the cell viability in a time-and dose-dependent manner (P<0.05), increased the percentage of apoptotic cells and the percentage of S-phase cells significantly, repressed the migration and invasion abilities (P<0.05), and decreased P21 expression at protein level (P<0.01). No influence on the expression of E-cadherin and vimentin in the BT549 cells was observed. CONCLUSION: AR antagonist HC-1119 decreases the viability, migration ability and invasion ability, enhances the apoptosis, and arrests the cell cycle distribution of TNBC BT549 cells. HC-1119 represses the viability of BT549 cells by down-regulating P21 expression, while the process of epithelial-mesenchymal transition is not involved in the inhibition of cell migration. 相似文献
18.
AIM: To investigate the effect of dihydroartemisinin (DHA) adjuvant treatment on enhancing the antitumor effect of 5-fluorouracil (5-FU) against gastric cancer. METHODS: The gastric cancer BGC-823 cells were divided into control group, DHA group, 5-FU group, 5-FU+DHA group and 5-FU+DHA+SIRT1 plasmid group. The viability of BGC-823 cells treated with DHA and 5-FU was measured by MTT assay. The expression of SIRT1 and NADPH oxidase, activation of caspase-9 and caspase-3, and phosphorylation of ASK1 and JNK in the BGC-823 cells treated with DHA and 5-FU were determined by Western blot. The production of ROS and the apoptosis of the BGC-823 cells treated with DHA and 5-FU were analyzed by flow cytometry. RESULTS: Dihydroartemisinin significantly inhibited the expression of SIRT1 and increased NADPH oxidase protein level (P<0.05). DHA increased the sensitivity of BGC-823 cells to 5-FU, thus decreasing the IC50 of 5-FU to the gastric cancer cells. However, transfection with SIRT1 plasmid decreased the cytotoxicity of DHA and 5-FU co-treatment to the BGC-823 cells. DHA promoted the production of ROS and phosphorylation of ASK1 and JNK induced by 5-FU in the BGC-823 cells (P<0.05). However, ROS scavenger N-acetylcysteine (NAC) or JNK specific inhibitor SP600125 inhibited the cell death and activation of caspase-9 and caspase-3 induced by DHA and 5-FU co-treatment (P<0.05). In addition, NAC significantly inhibited the phosphorylation of JNK in the BGC-823 cells co-treated with DHA and 5-FU. However, treatment with SP600125 did not influence the ROS production in the BGC-823 cells, indicating that JNK was the downstream target of ROS pathway. CONCLUSION: Combination of DHA with 5-FU induces caspase-dependent apoptosis in gastric cancer cells through the SIRT1/NADPH oxidase/ROS/JNK signaling pathway. 相似文献
19.
AIM: To investigate the anti-angiogenic effect of thymoquinone on human breast cancer and the possible mechanism. METHODS: The activities of human umbilical vein endothelial cells (HUVECs) and human breast cancer MCF-7 cells were assessed by CCK-8 assay. The apoptotic rate was measured by flow cytometry. Furthermore, the assay of capillary tube formation was used to observe the effect of thymoquinone on the tube formation of HUVECs. The protein levels of cleaved caspase-3, p-ERK and p-AKT were detected by Western blot. MCF-7 cells were subcutaneously injected into nude mice to establish breast xenograft tumors. After 3 weeks of implantation, the mice were randomized into control group and thymoquinone group. After the mice were sacrificed, immunohistochemistry was used to detect the expression of CD31 in the tumors, and the TUNEL test kit was used to explore cell apoptosis in the tumor tissues. RESULTS: Thymoquinone at concentrations of 20~80 nmol/L exerted no growth inhibitiory effect on MCF-7 cells. However, the cell viability of HUVECs were (66.1±8.3)％, (53.7±3.4)％ and (41.6±4.9)％ when the concentrations of thymoquinone were 20, 40 and 80 nmol/L, respectively. The apoptotic ratio of MCF-7 cells were (2.6±0.3)％, (2.4±0.3)％ and (4.6±0.4)％ and the apoptotic ratio of HUVECs were (21.5±3.7)％, (23.8±2.9)％ and (27.8±3.1)％ when the concentrations of thymoquinone were 20, 40 and 80 nmol/L, respectively. HUVECs were more sensitive to thymoquinone-induced apoptosis and inhibition in the cell activity than MCF-7 cells. Incubation of HUVECs with diffe-rent concentrations of thymoquinone (20, 40 and 80 nmol/L) for 1 h decreased their tube formation capacity (P<0.05). The protein level of cleaved caspase-3 was up-regulated, but the phosphorylation of AKT and ERK were inhibited in a concentration-dependent manner. The immunohistochemical analysis of CD31 showed significant difference of the integral absorbance between control group and thymoquinone group, and the TUNEL-positive cells in thymoquinone group was significantly more than that in control group. CONCLUSION: Thymoquinone has the anti-angiogenic effect on breast cancer both in vitro and in vivo, which may be related to the decreases in p-ERK and p-AKT. 相似文献
20.
AIM: To investigate the expression of CUG-binding protein 1 (CUGBP1) in breast cancer tissues, and to explore the effect of CUGBP1 gene silencing on the viability and invasion ability of human breast cancer MCF-7 cells. METHODS: A total of 96 cases of patients with breast cancer undergoing surgical treatment were selected in the Second Affiliated Hospital of Zhengzhou University from March 2015 to September 2017. Immunohistochemical staining was used to detect the protein expression of CUGBP1 in the breast cancer and adjacent tissues. MCF-7 cells were cultured and divided into CUGBP1 interference sequence group, control sequence group and blank group. Western blot was used to detect the protein expression of CUGBP1, Twist, E-cadherin and vimentin in the cells. The cell viability was measured by MTT assay. The cell invasion ability was detected by Transwell assay. RESULTS: The positive expression rate of CUGBP1 protein in the breast cancer tissues was higher than that in the adjacent tissues (χ2=28.900, P<0.001). The differences of CUGBP1 protein expression in the breast cancer tissues among TNM staging, histological grading and lymph node metastasis were statistically significant (P<0.05). The relative protein expression levels of CUGBP1, Twist and vimentin in CUGBP1 interference sequence group were lower than those in control sequence group and blank group, while the relative protein expression of E-cadherin was higher than that in control sequence group and blank group (P<0.05). The cell viability at 24 h, 48 h, 72 h and 96 h in CUGBP1 interference sequence group was lower than that in control sequence group and blank group (〖P<0.05). The invasive cells in CUGBP1 interference sequence group were less than those in control sequence group and blank group (P<0.05). CONCLUSION: CUGBP1 protein is highly expressed in the breast cancer tissues. Specific silencing of 〖STBX〗CUGBP1〖STBZ〗 gene expression in breast cancer MCF-7 cells effectively inhibits the cell viability and invasiveness, and its mechanism may be related to inhibiting the process of epithelial-mesenchymal transition. 相似文献