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目的 研究hCacyBP编码基因在胃癌多药耐药机制中的作用。方法 采用Northern杂交 ,检测SGC790 1细胞及SGC790 1/ADR细胞中hCacyBPmRNA表达水平的差异。将hCacyBPcDNA克隆到 pcDNA3.1中 ,构建反义核酸真核表达载体 pcDNA3.1/hCacyBP ,并转染耐阿霉素人胃癌细胞 ,用RT PCR检测转染细胞中hCacyBPmRNA水平的变化。用MTT比色法和FCM ,分别检测SGC790 1/ADR细胞、pcDNA3.1/hCacyBP 和 pcDNA3.1分别转染的SGC790 1/ADR细胞 ,对ADR的药物敏感性和胞内ADR的蓄积浓度。结果 Northern杂交证实 ,SGC790 1/ADR细胞中hCacyBPmRNA表达的水平显著高于SGC790 1细胞。成功地构建了pcDNA3.1/hCacyBP 。以 pcDNA3.1/hCacyBP 转染的SGC790 1/ADR细胞中hCacyBPmRNA的表达水平 ,显著低于空载体转染及未转染的SGC790 1/ADR细胞。MTT检测表明 ,转染pcDNA3.1/hCacyBP 的细胞 ,对ADR的药物敏性较空载体转染及未转染的SGC790 1/ADR细胞有所增高 ,生存率较后两者为低。FCM显示 ,转染 pcD NA3.1/hCacyBP 的SGC790 1/ADR细胞、转染 pcDNA3.1及未转染的SGC790 1/ADR细胞内ADR的蓄积浓度 ,分别为6 .72、5 .6 2和 5 .5 4。结论 hCacyBP碥码基因对胃癌细胞的MDR有一定的影响 ,CacyBP可能是一种胃癌细胞MDR中的重要分子
Objective To study the role of hCacyBP gene in gastric cancer multidrug resistance. Methods Northern hybridization was used to detect the difference of hCacyBP mRNA expression in SGC790 1 cells and SGC790 1/ADR cells. The hCacyBP cDNA was cloned into pcDNA3.1, and the antisense eukaryotic expression vector pcDNA3.1/hCacyBP was constructed and transfected with adriamycin-resistant human gastric cancer cells. The level of hCacyBP mRNA in the transfected cells was detected by RT PCR. MTT colorimetry and FCM were used to detect the sensitivity of ADR and the accumulation of intracellular ADR in SGC7901/ADR cells transfected with SGC7901/ADR cells, pcDNA3.1/hCacyBP and pcDNA3.1, respectively. Results Northern hybridization confirmed that the level of hCacyBP mRNA expression in SGC790 1/ADR cells was significantly higher than that of SGC790 1 cells. The pcDNA3.1/hCacyBP was successfully constructed. The expression of hCacyBP mRNA in SGC7901/ADR cells transfected with pcDNA3.1/hCacyBP was significantly lower than that of the empty vector transfected and untransfected SGC7901/ADR cells. MTT assay showed that the cells transfected with pcDNA3.1/hCacyBP were more sensitive to ADR than the empty vector transfected and untransfected SGC7901/ADR cells, and the survival rate was lower than the latter two. FCM showed that the accumulation of ADR in SGC790 1/ADR cells transfected with pcD NA3.1/hCacyBP, transfected with pcDNA3.1, and untransfected SGC790 1/ADR cells was 6.72, 5.62 and 5 .5 4. Conclusion The hCacyBP gene has a certain influence on MDR of gastric cancer cells. CacyBP may be an important molecule in gastric cancer cell MDR.