生存素反义核酸增加人结肠癌对泰素帝的化疗敏感性

目的探讨生存素(survivin)反义RNA真核表达质粒pcDNA3.0/survivin对泰素帝诱导人结肠癌多药耐药细胞株LOVO/Adr凋亡的影响。方法将已构建成功的survivin反义RNA真核表达质粒pcDNA3.0/survivin用脂质体瞬时转染LOVO/Adr细胞,以RT-PCR法检测质粒转染前后细胞survivinmRNA的变化,用MTT法和流式细胞术分别观察泰素帝对转染细胞的毒性作用和细胞凋亡变化。结果pcDNA3.0/survivin明显下调LOVO/Adr细胞survivinmRNA表达,MTT检测发现,泰素帝对转染pcDNA3.0/survivin、pcDNA3.0及未转染细胞抑制率分别为(37.3±2.9)%,(21.9±2.3)%和(21.1±1.9)%,前者与后两者间差异具有统计学意义(P<0.01);流式仪分析显示,各组细胞凋亡率分别为(28.7±1.7)%、(13.4±1.6)%与(14.3±1.8)%,前者与后两者间差异有统计学意义(P<0.01)。结论Survivin反义RNA真核表达质粒pcDNA3.0/survivin能下调LOVO/Adr细胞survivin基因表达,增加其对泰素帝的敏感性,为临床提高结肠癌疗效提供了一种新思路。

Antisense RNA targeting survivin enhances the chemosensitivity of LOVO/Adr cells to taxotere

OBJECTIVE: To investigate the ability of antisense RNA eukaryotic expression plasmid pcDNA3.0/survivin targeting survivin gene to inhibit survivin expression and enhance the sensitivity to taxotere in multidrug resistant colon carcinoma cell line LOVO/Adr. METHODS: The antisense RNA eukaryotic plasmid pcDNA3.0/survivin was transfected into LOVO/Adr cells by lipofectamine. The expression of survivin mRNA was measured using RT-PCR. After treated with taxotere, MTT assay and flow cytometry were used to evaluate the proliferation inhibition and apoptosis of LOVO/Adr cells. RESULTS: The expression of survivin mRNA in LOVO/Adr cells transfected with pcDNA3.0/survivin was down-regulated in a time- dependent manner. The inhibitory rate of taxotere (0.5 micromol/L) was (37.3 +/- 2.9)% in pcDNA3.0/survivin transfected cells, significantly higher than (21.9 +/- 2.3)% and (21.1 +/- 1.9)% in pcDNA3.0 transfected and untransfected control cells respectively (P< 0.01). The apoptosis rate of taxotere was (28.7 +/- 1.7)% in pcDNA3.0/survivin transfected cells,significantly higher than (13.4 +/- 1.6)% and (14.3 +/- 1.8)% in pcDNA3.0 transfected and untransfected cells respectively. CONCLUSION: The antisense RNA eukaryotic expression plasmid pcDNA3.0/survivin could down-regulate the expression of survivin gene and enhance the chemosensitivity of LOVO/Adr cells to taxotere, which may provide a novel therapy for colon carcinoma.