Anti-Sonic hedgehog blocking antibody enhances killing effect of PBMCs on cervical carcinoma HeLa cells     

SUN Yan  HUANG Li-xia  HUANG Wen-hao  HUANG Bin 《园艺学报》2011,27(12):2241-2246

AIM: To investigate the effect of anti-Sonic hedgehog(Shh) blocking antibody on the killing effect of peripheral blood mononuclear cells(PBMCs) on cervical carcinoma HeLa cells. METHODS: The expression levels of Shh and Shh signaling molecules in HeLa cells were detected by immunocytochemistry and RT-PCR. PBMCs from health peoples were isolated by the method of Ficoll density gradient centrifugation, and then co-cultured with HeLa cells in vitro. The expression of CD3, CD69 and CD71 was assayed by flew cytometry. The concentrations of IFN-γ, IL-10 and IL-4 in culture supernatants were detected by ELISA. The killing effect of PBMCs on HeLa cells was observed under microscope. RESULTS: Shh and Shh signaling molecules were expressed in HeLa cells. The level of Shh expression didn't change significantly in the 6th passage of HeLa cells. CD3⁺ cells were increased in the co-culture system. The expression of CD69 and CD 71, and the secretion of IFN-γ were increased, while the secretion of IL-10 was decreased in the co-culture system treated with anti-Shh blocking antibody. Anti-Shh blocking antibody has no effect on the secretion of IL-4. The killing effect of PBMCs on HeLa cells was strengthened by anti-Shh blocking antibody. CONCLUSION: Anti-Shh blocking antibody promotes the activation of PBMCs and enhances the killing effect of PBMCs on cervical carcinoma HeLa cells.  相似文献   

Inhibition of Ang-2 and RGS-5 expression by nanogold results in normalization of vascellum in hepatic tumor     

FU Yue-wu  PAN Yun-long  QIN Li  SUN Li  LIU Ying-mei 《园艺学报》2011,27(12):2247-2252

AIM: To observe the vascular normalization effect of nanogold on hepatic tumor by inhibiting angiopoietin-2 (Ang-2), regulator of G-protein signaling-5 (RGS-5) during certain time window. METHODS: H22 cells, the hepatocellular cancer cell line, were subcutaneously injected into the right armpits of 48 BALB/c nude mice. When the size of transplanted tumor reached 3~4 mm, the mice were divided into 2 groups: 36 mice in experiment group and 12 mice in control group. The mice in experimental group underwent injection of nanogold into the tumor once a day, and the mice in control group were injected with normal saline. After continuous treatment with nanogold for 3 days, 7 days and 11 days, the mice were sacrificed, the liver tumors were taken out to measure the size and weight. The expression of Ang-1,Ang-2 and RGS-5 in the tumor was detected by the method of immunohistochemical staining. The normalizing shapes of tumor vessels and the pericytes were observed under electronic microscope. RESULTS: With nanogold treatment for 3 days, 7 days and 11 days, the positive rates of Ang-1 were 16.7％, 50.0% and 16.7%, respectively. The positive rates of Ang-2 were 33.3%, 16.7% and 41.7%, respectively. The expression of Ang-1 in experiment group was higher than that in control group, especially at the 7th day in experiment group. The expression of Ang-2 in experiment group was lower than that in control group (58.3%), especially at the 7th day in experiment group. With nanogold treatment for 3 days, 7 days and 11 days, the positive rates of RGS-5 were 33.3%, 16.7% and 50.0%, respectively. The immature pericyte coverage indexes (IMPI) were 19.6%±4.3%, 32.5%±7.9% and 41.2%±9.1% respectively. At the 7th day in experiment group, the positive rates of RGS-5 and IMPI were lower than those in other experiment groups and control group. After treated with nanogold for 7 days, the pericytes in the parietal wall of the blood vessels in the tumor showed the tendency to grow normally in morphology and were completely covered by endothelial cells. However, the pericytes in the parietal wall of the blood vessels in control group showed differences in size, impaired integrity and only a few of the pericytes covered by endothelial cells. CONCLUSION: During the time window of nangold treatment for 7 days, the chemical can normalize the blood vessels in liver cancer by inhibiting the expression of Ang-2 and RGS-5.  相似文献   

Effect of prostate-specific membrane antigen on phospho-ERK,growth and migration of prostate cancer LNCaP cells     

GUO Zheng-hui  HUANG Hai  DU Tao  XU Ke-wei  CAO Yi  CHEN Jie-qing  DONG Wen  YAO You-sheng  LIN Tian-xin  XIE Wen-lian  JIANG Chun  HAN Jin-li  HUANG Jian 《园艺学报》2011,27(6):1090-1096

AIM: To study the expression of prostate-specific membrane antigen (PSMA) on the level of phospho-ERK, cell growth and migration of prostate cancer LNCaP cells.METHODS: The method of silencing PSMA was established by lentivirus-mediated RNAi in our early experiment. The cells were divided into 3 groups.In experimental group, the expression of PSMA in LNCaP cells was stably blocked by lentivirus-mediated RNAi. In negative control group, the cells were transfected with lentivirus-mediated control RNAi (without any interference to PSMA).The normal LNCaP cells served as blank control. The cells in these 3 groups were cultured in both 2 environments: normal medium and medium with PD98059 (an inhibitor of ERK phosphorylation). The phospho-ERK was detected by Western blotting and immunocytochemistry. Furthermore, the growth and migration of the cells were evaluated by MTT and transwell assays,respectively.RESULTS: In normal medium, the expression of phospho-ERK was attenuated in experimental group (P<0.05) and the quantity of "positive" cells was less than those in other 2 groups (P<0.05). Furthermore, the growth curves of the cells showed that the growth ability in experimental group was significantly decreased (P<0.05, after 48 h) and the migration ability in experimental group was reduced (P<0.05). In the inhibitory medium, the cells in all 3 groups expressed phospho-ERK at a lower level. Moreover, the abilities of growth and migration in these 3 groups were poorly displayed. These inhibitory effects on phosphorylation of ERK were similar to the cells in experimental group cultured in normal medium.CONCLUSION: PSMA may play a role in up-regulation of phospho-ERK and it may take an advantage in growth and migration of prostate cancer LNCaP cells.  相似文献   

Bufalin activates chloride channels in poorly differentiated nasopharyngeal carcinoma cells     

LUO Hai-bing  LI Hua-rong  LIU Shan-wen  PENG Shuang  LIU Zhen- feng  YE Wen-cai  ZHANG Dong-mei  CHEN Li-xin  WANG Li-wei 《园艺学报》2011,27(4):672-676

AIM: To investigate the activation of chloride channels induced by bufalin and the properties of the channels in poorly differentiated nasopharyngeal carcinoma (CNE-2Z) cells. METHODS: The technique of whole-cell patch clamp was used to record the chloride currents and to analyze the characteristics of the currents in CNE-2Z cells.RESULTS: A chloride current was slowly activated by extracellular application of bufalin (1 μmol/L). The activation of the current was slower than that of the volume-activated chloride current, with an activation latency of(12.1±6.4) min. The reversal potential of the current was close to the calculated Cl^- equilibrium potential (E_Cl =-0.9 mV). The chloride current was outward-rectified and did not show significant time-dependent or voltage-dependent inactivation. The chloride channel blocker tamoxifen completely inhibited the outward and inward currents. The current was also completely inhibited by extra-cellular application of 47% hypertonic solution. CONCLUSION: Bufalin activates chloride channels and induces a chloride current in CNE-2Z cells. Compared with the volume-activated chloride current in CNE-2Z cells, the activation latency of the bufalin-induced current is longer and the outward rectification is more obvious.  相似文献   

ULTRASONOGRAPHY OF THE CANINE, FELINE AND EQUINE TONGUE: NORMAL FINDINGS AND CASE HISTORY REPORTS   总被引：3，自引：0，他引：3  

Mauricio  Solano  Med. Vet.  Dominique G.  Penninck  DVM 《Veterinary radiology & ultrasound》1996,37(3):206-213

A submental approach using a 5, 7.5 and 10 MHz mechanical sectorial transducers was used to evaluate the ultrasonographic appearance of the normal and abnormal canine, feline and equine tongue. Seven normal animals were used to characterize the in vivo normal ultrasonographic appearance of the oropharynx. Consistently seen anatomical landmarks included the lingual parenchyma, lingual artery, soft and hard palate interfaces, mandible, basihyoid bone and sublingual musculature. Seven fresh canine, feline and equine cadavers were dissected to confirm the ultrasonographic findings. Seven animals with signs of ptyalism, dysphagia, halitosis and visible lingual lesions were also studied. A foreign body and abscesses secondary to a penetrating wound were detected in a horse. Ultrasound guidance aided surgical dissection, removal of the foreign body and intraoperative drainage of the abscesses. Ultrasound provided valuable non-invasive documentation of the architecture, size and margins of six lingual neoplasms. Ultrasonographic monitoring of the response to the treatment of lingual lesions may be beneficial in the management of patients.  相似文献   

Identification of specific serum biomarkers in gastric adenocarcinoma patients     

WANG Wei-ping  LIU Chi-bo 《园艺学报》2012,28(2):274-280

AIM: To screen the possible serum biomarkers for the diagnosis of gastric adenocarcinoma. METHODS: The surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) was used to screen the serum samples from 109 cases of gastric adenocarcinoma and 106 control subjects (60 healthy subjects, 30 patients with chronic superficial gastritis and 16 cases of chronic atrophic gastritis). The differentially-expressed protein peaks were selected and isolated with high-performance liquid chromatography (HPLC), and processed with enzyme before liquid chromatography-mass spectrometry tandem mass spectrometry (LC-MS/MS) analysis. The data mining was performed with software Xcalibur program component Bioworks 3.2. RESULTS: Three differentially-expressed protein peaks were selected as potential serum biomarkers of gastric adenocarcinoma patients.The m/z peak at 5 906.5 showed the increase (8.53±4.33 in cancer group, and 0.88±0.31 in control group). The m/z peaks at 6 635.7 and 8 716.3 showed the decrease (6.54±2.44 and 0.93 ± 0.29, respectively, in cancer group and 17.56±4.43 and 2.16±0.98, respectively, in control group, P<0.01). The 3 m/z peaks were identified as fibrinogen α-chain, apolipoprotein A-II and apolipoprotein CI,respectively. The combined use of the 3 biomarkers distinguished the samples in the cancer patients out of the controls at a sensitivity of 93.85% (61/65) and a specificity of 94.34% (50/53). CONCLUSION: The fibrinogen α-chain, apolipoprotein A-II and apolipoprotein CI identified as potential markers for gastric adenocarcinoma show diagnostic values in clinical application.  相似文献   

Clinical significance and prognostic value of extracellular signal-regulated kinases in epithelial ovarian cancer     

NI Juan  CHEN Ya-xia  LU Hong-xian  CHENG Bei  WAN Xiao-yun  XIE Xing 《园艺学报》2012,(1):65-69

AIM: To investigate the expression of phosphorylated extracellular signal-regulated kinase (p-ERK) in epithelial ovarian cancer and to analyze the correlation between the expression of p-ERK and prognosis of epithelial ovarian cancer patients. METHODS: The samples of normal ovarian tissues, benign ovarian tissues and ovarian carcinoma were collected. The expression of p-ERK in each sample was detected by Western blotting. Assemble clinical data and survival were analyzed with Kaplan-Meier method and Cox regression. RESULTS: The expression level of p-ERK in ovarian carcinoma was significantly higher than that in benign ovarian tissues and in normal ovarian tissues. The overexpression of p-ERK protein was significantly higher in stage Ⅲ/Ⅳ disease than that in stage I/II disease. No difference of p-ERK expression level between the benign ovarian tissues and the normal ovarian tissues was observed. The overexpression of p-ERK was found more often in poorly-differentiated ovarian cancer. Compared with serous carcinoma of ovary, the expression level of p-ERK was similar in undifferentiated adenocarcinoma. The expression level of p-ERK was low in endometrioid carcinoma of ovary and mucinous carcinoma of ovary. The expression level of p-ERK was negatively correlated with the survival time. Increased p-ERK expression was strongly correlated to the poor survival. Shorter survival time was significantly associated with p-ERK expression, residual tumor, stage, grade and chemotherapy course. Shorter survival time was not significantly correlated with tumorous pathology. CONCLUSION: p-ERK is a key regulation kinase involved in epithelial ovarian cancer. The expression level of p-ERK seems to have a prognosis value in epithelial ovarian cancer.  相似文献   

Expression of ERK5 in colorectal cancer tissues and its correlation with distant metastasis in colorectal cancer patients     

HU Bang  XIE Shang-kui  HUANG Mei-jin  JIANG Li  PENG Hui  WANG Jian-ping  REN Dong-lin  WANG Lei 《园艺学报》2012,28(7):1213-1218

AIM: To investigate the expression of extracellular signal-regulated kinase 5 (ERK5) in primary colorectal cancer (CRC) and adjacent normal mucosa, and to analyze the relationship between ERK5 expression and clinicopathological parameters for exploring the functions of ERK5 in the occurrence and development of CRC. METHODS: The expression of ERK5 in carcinoma tissues and normal mucosa was examined by a set of tissue microarrays and the method of immunohistochemistry. The potential relationship between ERK5 expression and clinicopathological features was also analyzed. RESULTS: ERK5 expression was significantly higher in CRC tissues (134/338, 39.6%) than that in normal tissues (21/80, 26.2%; P<0.05). Overexpression of ERK5 in CRC tissues was significantly correlated with distant metastasis (P<0.05). However, no correlation between ERK5 expression and age at surgery, sex, tumor location, the depth of invasion, lymph node metastasis, TNM staging or differentiation grade was found (P>0.05). According to the Kaplan-Meier analysis, there is no significant difference in 5-year overall survival between the patients with ERK5 expression at high level and at low level. CONCLUSION: ERK5 protein is highly expressed in CRC with distant metastasis. This may be a promotive factor in the process of distant metastasis.  相似文献   

Effects of ATP-competitive GSK-3 inhibitor on multidrug resistance of human esophageal squamous-cell carcinoma cells     

GAN Si-yuan  ZHONG Xue-yun  XIE Si-ming  LUO Feng 《园艺学报》2012,28(7):1219-1223

AIM: To study the changes and correlations of β-catenin, multidrug resistance-associated protein 2 (MRP2), P-glycoprotein (P-gp), Bcl-2 and the free ATP level in esophageal squamous-cell carcinoma (ESCC) cell line EC-109 induced by ATP-competitive glycogen synthase kinase(GSK-3) inhibitor 6-bromoindirubin-3'-oxime (BIO). METHODS: The methods of flow cytometry, immunofluorescence, cytochemistry and ATP assay were used to study the expression levels of MRP2, P-gp, β-catenin and Bcl-2, the efflux capability of ATP-binding cassette (ABC) transporters, and the free ATP level in ESCC cells. RESULTS: After induced by BIO, up-regulation of β-catenin and Bcl-2 expression in cytoplasm and accumulation of β-catenin in nucleus were found in ESCC cells. The expression of MRP2 was up-regulated in cytoplasm and cell membrane. On the contrary, the expression of P-gp was down-regulated in cytoplasm and cell membrane. The free ATP level and the efflux capability of ABC transporters increased in ESCC cells. CONCLUSION: The multidrug resistance of ESCC cells in enhanced by BIO treatment.  相似文献   

Expression of EPCAM,CD44 and CD24 in gastric cancer tissues and its clinical significance     

YANG Shi-bin  XIAO Long-bin  XU Feng-feng  WU Wen-hui  LI Shou-zhi  LI Ming-zhe 《园艺学报》2012,28(7):1224-1229

AIM: To evaluate the relationship between epithelial cell adhesion molecule (EPCAM)/cluster of differentiation 44 (CD44)/cluster of differentiation 24 (CD24) expression and the clinicopathological characteristics/prognosis in 95 gastric cancer patients. METHODS: The expression levels of EPCAM, CD44 and CD24 were detected using the two-step method of immunohistochemistry in 95 gastric cancer patients who underwent surgical excision and were pathologically diagnosed as gastric cancer. RESULTS: There were 56 EPCAM-positive patients (58.95%), 41 CD44-positive patients (43.16%) and 56 CD24-positive patients (58.95%). Thirty patients were both EPCAM and CD44 positive (31.58%), 45 patients were both EPCAM and CD24 positive (47.37%), 32 patients were both CD44 and CD24 positive (33.68%), and 25 patients were EPCAM, CD44 and CD24 positive (26.32%). EPCAM expression was correlated with age, depth of tumor infiltration and WHO histological classification. CD44 expression was correlated with BORRMANN and WHO histological classification as well as CEA value. CD24 expression was correlated with the depth of infiltration, location of the tumor, WHO histological classification and viscera invasion. All positive expression of EPCAM, CD44 and CD24 was correlated with the depth of infiltration, location of the tumor and WHO histological classification (P<0.05). The difference of survival rate between EPCAM positive group and negative group was observed, and the CD44 positive group and negative group had the same result (P<0.05 and P<0.01, respectively). The difference of survival rate between EPCAM⁺CD44⁺CD24⁺ group and EPCAM^-CD44^-CD24^- group was statistically significant (P<0.05). The difference of survival rate between EPCAM^-CD44⁺CD24⁺ group and EPCAM^-CD44^-CD24^- group was also significant (P<0.05). CONCLUSION: The positive rates of EPCAM, CD44 and CD24 expression are high in gastric cancer tissues and these 3 proteins can be used as primary screening targets.  相似文献