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目的筛选出结肠癌细胞侵袭相关研究的最佳培养基。方法在RPMI 1640、DMEM、DMEM-F12培养基中对人结直肠癌细胞株HCT-116、LOVO、CACO-2进行常规培养,采用集落形成实验检测肿瘤细胞增殖能力,免疫荧光检测上皮型钙黏素(E-cadherin)、波形蛋白(vimentin)、Nm23-H1、p53蛋白表达,RT-PCR检测Nm23-H1、p53、蜗牛基因(snail-1)mRNA表达。结果与RPMI-1640、DMEM培养组相比,DMEM-F12培养组三种人结直肠癌细胞克隆集落形成率、vimentin、p53蛋白、snail-1和p53mRNA表达均明显增加(P<0.05),而E-cadherin、Nm23-H1蛋白表达和Nm23-H1mRNA表达降低(P<0.05);RPMI 1640培养组与DMEM培养组间各观察指标差异均无统计学意义(P>0.05)。结论 DMEM-F12培养基相对DMEM、RPMI 1640培养基更适用于结肠癌细胞侵袭性能相关的基础实验研究。
Objective To screen out the best culture medium for colon cancer cell invasion related research. Methods Human colorectal cancer cell lines HCT-116, LOVO and CACO-2 were cultured in RPMI 1640, DMEM and DMEM-F12 medium. The colony formation assay was used to detect the proliferation of tumor cells. E-cadherin, vimentin, Nm23-H1 and p53 were detected by RT-PCR. The expressions of Nm23-H1, p53 and snail-1 mRNA were detected by RT- Results Compared with RPMI-1640 and DMEM group, the colony formation rate, the expression of vimentin, p53 protein, snail-1 and p53 mRNA in three human colorectal cancer cell lines were significantly increased (P <0.05) The expression of E-cadherin, Nm23-H1 protein and Nm23-H1 mRNA decreased (P <0.05). There was no significant difference between the RPMI 1640 culture group and DMEM culture group (P> 0.05). Conclusion DMEM-F12 medium is more suitable for the basic experimental research of colon cancer cell invasion than DMEM and RPMI 1640 medium.