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1.
AIM: To observe the effect of antisense locked nucleic acid (anti-LNA) blocking the translation initiation region of c-myc exon 2 on the viability and apoptosis of hepatocellular carcinoma cells.METHODS: The anti-LNA that was complementary to the translation initiation region of c-myc exon 2 was designed, synthesized, and introduced into the HepG2 cells by cationic liposome-mediated transfection. The mRNA and protein levels of c-Myc in the cells were determined by RT-PCR and Western blot. The change of cell apoptosis was analyzed by flow cytometry, and the toxicity of anti-LNA to the cells was detected by MTT assay.RESULTS: Five days after transfection, the mRNA level of c-Myc in anti-LNA group was 0.335±0.016, and the protein level was 0.448±0.037, significantly lower than those in control group (both P<0.05). The ratio of apoptotic cells in anti-LNA group was 32%±6%, which was higher than that in control group (P<0.05).CONCLUSION: Antisense locked nucleic acid targeting at the translation initiation region of c-myc exon 2 shows strong inhibitory effects on the apoptosis of hepatocellular carcinoma cells. 相似文献
2.
探讨利用小鼠制备抗人c—myc蛋白多克隆抗体的方法。利用含PET28a／c—myc质粒的EcoliBL21,对c—myc蛋白进行原核表达,并进行纯化。WesternBlot结果只出现一条62KDa大小的特异性目的蛋白条带。用原核表达的c—myc蛋白抗原免疫5只Balb／c小鼠,最终免疫后的第7d采血,并对其血清效价进行ELISA（Enzyme—linked immunosorbnent assay）测定,抗血清达1：640000。本实验结果为c—myc单克隆抗体制备和肿瘤诊断试剂盒的研发提供基础实验数据。 相似文献
3.
利用细胞培养、原位杂交、免疫组化、图像分析等技术研究缺氧与肉鸡肺动脉平滑肌细胞癌基因c-fos和c-myc表达的关系,进一步阐明缺氧与以肺动脉增殖重塑为特征的肉鸡腹水综合征的关系。结果显示,缺氧显著引起肺动脉平滑肌细胞内癌基因c-fos和c-mycmRNA的表达(c-fosmRNA:常氧组为144.6±20.2,缺氧组为198.1±32.8,P<0.01;c-mycmRNA:常氧组为125.4±18.8,缺氧组为167.1±22.4,P<0.01)。缺氧显著引起肺动脉平滑肌细胞内癌基因c-fos和c-myc蛋白的表达(c-fos蛋白:常氧组为150.9±33.2,缺氧组为225.9±37.0,P<0.01;c-myc蛋白:常氧组为162.1±28.5,缺氧组为228.8±33.4,P<0.01)。结果表明缺氧能够明显诱发肉鸡肺动脉平滑肌细胞内癌基因c-fos、c-myc的转录和表达,是启动肺动脉平滑肌细胞增殖的重要原因。本研究阐明缺氧是以肺动脉重塑为特征的肉鸡腹水综合征的主要诱因。 相似文献
4.
AIM: The cancer biology has showed that overexpression of oncogenes is responsible for the progression of human malignancies,antisense technology can block a certain gene expression.Caffeine has enhancement effect on chemotherapy of osteosarcoma cells to cisplatin,we constructed the recombinant adenovirus (Ad-Asc-myc) encoding antisense c-myc fragment and investigated its effect on the in vitro sensitivity of osteosarcoma MG-63 cells to cisplatin.METHODS: The recombinant adenovirus (Ad-Asc-myc) encoding antisense c-myc fragment was constructed by cloning c-myc cDNA of about 750 base pairs in a reverse direction into adenovirus vector.Ad-Asc-myc and caffeine was used respectively or together to co-operate with cisplatin to treat the osteosarcoma MG-63 cells in vitro,and Western blotting,MTT,flow cytometry (FCM),electron microscope were used to evaluate expression of c-Myc protein,tumor cell proliferation in vitro,apoptosis and cell cycle analysis.RESULTS: Ad-Asc-myc was obtained with the titer of 2×1012 pfu/L.Ad-Asc-myc down-regulated the expression of c-Myc protein,Ad-Asc-myc or caffeine enhanced the effects of 2.0,5.0 mg/L cisplatin on MG-63 cells.Moreover,Ad-Asc-myc combined with caffeine significantly enhanced this effects,not only on cisplatin-induced apoptosis,but also on tumor cells proliferation in vitro.The expression of bcl-2 was downregulated,bax were upregulated,while there was no change in the expression of E2F-1.FCM analysis showed that cisplatin treatment induced a block in S phase,and caffeine reversed this block and speeded up the progression of cells out of the S phase.Ad-Asc-myc induced obvious G2/M phase arrest in transfected cells.CONCLUSION: Ad-Asc-myc combined with caffeine may enhance apoptosis-induced and chemotherapy effects of osteosarcoma MG-63 cells to cisplatin. 相似文献
5.
目的观察澳洲茄胺盐酸盐对HeLa细胞c-myc，H—ras和hTRT基因表达的影响．方法用鼠抗人c-mycMcAb，H-ras McAb和hTRTPcAb作为一抗，生物素化羊抗鼠IgG作为二抗，用免疫组化方法测定澳洲茄胺盐酸盐对HeLa细胞c-myc，H—ms和hTRT基因的表达．结果澳洲茄胺盐酸盐处理的HeLa细胞与对照组相比，c-myc，H-ras和hTRT基因的表达均明显降低（P〈0．05）．结论澳洲茄胺盐酸盐下调HeLa细胞c-myc和H-ras基因的表达，抑制细胞增殖，诱导分化；澳洲茄胺盐酸盐下调HeLa细胞hTRT基因的表达，抑制端粒酶活性，抑制细胞生长． 相似文献
6.
c-myc和c-fos在隆肛蛙与北方山溪鲵卵巢中的表达 总被引：1，自引：0，他引：1
用免疫细胞化学方法,对隆肛蛙与北方山溪鲵不同发育时期卵泡c- myc和c- fos的表达产物进行了定位检测。结果表明,Myc在2种动物 - 期卵母细胞和滤泡细胞中表达较强,以后逐渐减弱;Fos在2种动物卵泡发育初期表达很弱,在 - 期卵泡中表达增强。上述结果说明,c- myc在2种动物卵母细胞发育初期起重要作用,而c- fos在发育中后期起重要的调控作用,2种原癌基因可能通过调节类固醇激素的合成与分泌来调控卵母细胞的发育。 相似文献
7.
AIM: To observe the proliferation and apoptosis of ovarian cancer cells by silencing the expression of human pituitary tumor-transforming gene 1 ( hPTTG1 ) using RNA interference technique.METHODS: The chemically synthesized siRNA targeting hPTTG1 was transfected into ovarian cancer cell line A2780 in vitro. The expression levels of hPTTG1 and c-myc were examined by RT-PCR and Western blotting. Cell proliferation was measured by MTT colorimetric assay and -TdR incorporation test. Cell apoptosis was detected by flow cytometry with annexin V/PI and TUNEL labeling.RESULTS: The expression of hPTTG1 at mRNA and protein levels was inhibited after transfection of hPTTG1 siRNA. The inhibitory efficiency was 70.5%±3.9% and 63.8%±4.5%, respectively. The absorbance began to decrease 24 h after transfection of hPTTG1 siRNA,and the highest inhibitory rate was 42.9%±5.2% at 48 h post-transfection. Radioactive incorporation of -TdR in hPTTG1 siRNA group was lower than that in normal and negative groups. The survival rate declined while the apoptotic rate and necrotic rate increased in hPTTG1 siRNA group. Apoptotic index in hPTTG1 siRNA group was higher than that in normal and negative groups. The expression of c-myc at mRNA and protein levels was down-regulated.CONCLUSION: Cell proliferation is inhibited and cell apoptosis is induced by hPTTG1 siRNA through down-regulating the expression of c-myc. hPTTG1 can be regarded as a candidate gene for ovarian cancer gene therapy. 相似文献
8.
为了探索c-myc蛋白在羊驼皮肤中的定位及表达情况,以不同毛色的成年羊驼为研究对象,采用免疫组织化学方法检测c-myc蛋白在羊驼皮肤组织中的表达和定位.结果显示,c-myc在羊驼皮肤毛囊毛球部细胞中呈阳性表达,根据光密度值分析得出c-myc在棕色羊驼毛囊中显著表达,其相对表达量是白色羊驼的5.51倍.研究表明c-myc可能参与羊驼毛色形成. 相似文献
9.
以含有 1.6 kb的小鼠乳清酸蛋白 (WAP)上游调序列的 p CAT- WAP和含有人 c- myc c DNA的 p WM为原始质粒 ,构建了 WAP启动子调控下的 c- myc乳腺定位表达载体 p WCS。载体用 Bgl Bam H 酶切 ,回收 3.5 kb的基因片段 WAP- c- myc- SV40 Poly A,通过显微注射方法导入 C5 7BL/ 6 J× DBA/ 2 JF1 代小鼠受精卵的雄原核内。共注射10 0 0枚卵 ,将存活的约 6 0 0枚卵分别移植至 2 9只假孕母鼠输卵管内 ,获得仔鼠 45只。PCR检测 ,阳性鼠 9只 ,South-ern blot检测 ,阳性鼠 3只 ,其中 1只母鼠 ,2只公鼠 ,在饲养过程中 ,1只母鼠意外死亡。对 2只转基因公鼠的 F1代PCR检测表明 ,仅 1只公鼠具有遗传性 ,在所生的 2 7只 F1代小鼠中有 13只为 PCR阳性。 相似文献
10.
为探讨内毒素（ET）对大鼠肝脏c—myc基因的表达影响及阳离子（cA）对该过程的保护效应,本试验将72只清洁级SD大鼠随机分为对照组、内毒素组和阳离子A保护组,3组经相应处理后分别在3、4、8、12h采集肝脏作为样本。采用实时荧光定量RT—PCR检测法和免疫组化法分析,结果显示：在mRNA水平和蛋白水平上,ET可上调C—myc基因的表达,从而促进肝细胞凋i-,最终导致肝细胞损伤,CA则能明显下调c—myc基因的表达,从而抑制肝细胞凋亡,对ET诱导的肝损伤具有明显的保护效应。 相似文献