BCL-3基因沉默对BCL-3高表达的人大肠癌RKO细胞增殖和药物敏感性的影响

Effects of BCL-3 gene silencing on the proliferation and drug sensitivity of human colorectal cancer cell line RKO with high expression of BCL-3

目的:构建针对人BCL-3基因的RNA干扰(RNA interference,RNAi)慢病毒载体,观察其对大肠癌细胞株中BCL-3基因的沉默效应,以及基因沉默对细胞生物学行为及药物敏感性的影响。方法:运用RT-PCR及Western blotting方法检测5株大肠癌细胞株中BCL-3的表达状况;构建人BCL-3基因的RNAi慢病毒载体,并转染BCL-3高表达的大肠癌细胞株,运用Western blotting方法鉴定其对BCL-3基因的沉默效果;BCL-3基因沉默后运用细胞增殖实验及软琼脂克隆形成实验检测其对细胞增殖能力的影响,运用MTT方法检测大肠癌细胞株对化疗药物敏感性的变化。结果:BCL-3高表达于大肠癌细胞株RKO中;成功构建了BCL-3 RNAi慢病毒载体,Western blotting实验显示其可抑制RKO细胞中BCL-3蛋白的表达;抑制BCL-3的表达后,增殖实验表明RKO细胞增殖能力下降,软琼脂克隆形成实验表明RKO细胞的克隆形成率下降;MTT实验表明BCL-3表达抑制后奥沙利铂对RKO细胞的半数抑制浓度(median inhibitory concentration,IC50)显著降低。结论:BCL-3表达抑制后,BCL-3高表达的大肠癌细胞株RKO的增殖能力下降,并且其对奥沙利铂的敏感性增强。

AIM： To construct lentiviral vectors for RNA interference （RNAi） of BCL-3 gene, and to detect the changes of biological behaviors and drug sensitivity of coloreetal cancer cells after BCL-3 gene silencing. METHODS： The expression of BCL-3 in five human colorectal cancer cell lines was detected by RT-PCR and Western blotting. Lentivi- ral vectors for RNAi of BCL-3 gene were constructed and transfected into the human colorectal cancer cell line with high ex- pression of BCL-3, and then the silencing effect was detected by Western blotting. After BCL-3 gene silencing, the change of cell proliferation was detected by MTY assay and soft agar colony formation assay, and the change of drug sensitivity was detected by MTY assay. RESULTS： BCL-3 was highly expressed in human colorectal cancer cell line RKO. Lentiviral vectors for RNAi of BCL-3 gene were successfully constructed, and Western blotting showed that BCL-3-shRNA2 could efficiently inhibit the expression of BCL-3 protein in RKO cells. After BCL-3 gene silencing, the proliferation ability and colony formation rate of RKO cells were decreased, and the median inhibitory concentration of oxaliplatin for RKO cells also de- creased significantly. CONCLUSION： Inhibition of BCL-3 gene expression decreases the proliferation ability of human colorectal cell line RKO with high expression of BCL-3, and enhances the sensitivity of RKO ceils to oxaliplatin. [