反义人钙周期素结合蛋白编码基因转染对胃癌细胞多药耐药性的影响

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SGC7901细胞及SGC7901／ADR细胞中hCacyBP mRNA表达水平的差异。将hCacyBP cDNA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hCacyBP mRNA水平的变化。用MTT比色法和FCM，分别检测SGC7901／ADR细胞，pcDNA3.1/hCacyBP-和pcDNA3.1分别转染的SGC7901／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，SGC7901／ADR细胞中hCacyBP mRNA表达的水平显著高于SGC7901细胞。成功地构建了pcDNA3.1/hCacyBP-。以pcDNA3.1/hCacyBP-转染的SGC791／ADR细胞中hCacyBP mRNA的表达水平，显著低于空载体转染及未转染的SGC791／ADR细胞。MTT检测表明，转染pcDNA3.1/hCacyBP-细胞，对ADR的药物敏性较空载体转染及未转染的SGC7901／ADR细胞有所增高，生存率较后两者为低。FCM显示，转染pcDNA3.1/hCacyBP-的SGC7901／ADR细胞，转染pcDNA3.1及未转染的SGC7901／ADR细胞内ADR的蓄积浓度，分别为6．72，5．62和5．54。结论 hCacyBP编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。

The effects of transfection with gene encoding antise human calcyclin binding protein on gastric cancer cell MDR

Aim To research a role of hCacyBP gene in gastric cancer cell MDR mechanism. Mehods Northern blot was used to detect the difference of hCacyBP mRNA expression levels between SGC7901 and SGC7901/ADR cells. HCacyBP antisese nucleic acid eukaryotic expression vector pcDNA3.1/hCacyBP was constructed by cloning hCacyBP cDNA into vector pcDNA3.1, and then SGC7901/ADR cell was transfected via mediation of lipofectamine2000 . RT PCR was used to detect the hCacyBP mRNA expression level in the transtected cells. MTT colormetry was used to detect the sensitivity of SGC7901/ADR cells and SGC7901/ADR cells to ADR. FCM was used to detect the accumulation concentration of ADR in the cells mentioned above. Results Northern blot analysis showed that the hCacyBP mRNA expression level in SGC7901/ADR cells was higher than that in SGC7901 cells. pcDNA3.1/hCacyBP was successfully constructed. The hCacyBP mRNA expression level in SGC7901/ADR cells transfected by pcDNA3.1/hCacyBP was remarkably lower than those in SGC7901 cells transfected by pcDNA3.1 and SGC7901/ADR cells. Detection of MTT colorimetry analysis showed that the sensitivity of SGC7901/ADR cells transfected by pcDNA3.1/hCacyBP to ADR was higher than that of other two cells, but its survival rate was lower than the those of two cells. FCM showed that the accumulated concentration of ADR in the SGC7901/ADR cells transfected by pcDNA3.1/hCacyBP and pcDNA3.1, respectively and in SGC7901/ADR cells was 6.72, 5.62 and 5.54, respectively. Conclusion HCacyBP gene can affect the MDR of gastric cancer cells. CacyBP maybe an important molecule leading to MDR of gastric cancer cells.