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研究鬼臼毒素衍生物CIP-36对耐药人口腔鳞状上皮癌细胞KBV200的抗肿瘤活性及其作用机制。采用MTT法观察CIP-36对多种肿瘤细胞和正常细胞的体外抑制作用以及对KB和KBV200细胞生长曲线的作用,Hoechst荧光染色进行细胞凋亡检测,RT-PCR检测CIP-36对KB及KBV200细胞p53、p21、caspase-3、bax、mdr-1和bcl-2的mRNA表达的影响,免疫组织化学检测观察CIP-36对KBV200细胞P-gp表达的影响。结果表明,CIP-36对多种肿瘤细胞均有较好的抑制作用,且对耐药株细胞均有明显抑制作用。荧光染色结果显示,CIP-36可诱导KBV200细胞凋亡。同时CIP-36可剂量依赖性地增加KBV200及KB细胞的p53、p21、caspase-3及bax的mRNA表达,同时降低mdr-1和bcl-2的mRNA表达,与对照组比较差异均有统计学意义。免疫组织化学检测结果显示,CIP-36显著降低KBV200的P-gp表达。提示CIP-36可能通过影响多个与肿瘤耐药相关基因和蛋白的表达来克服肿瘤细胞株的多药耐药性。
To study the antitumor activity of podophyllotoxin derivative CIP-36 on human oral squamous cell carcinoma KBV200 cells and its mechanism. The inhibitory effect of CIP-36 on various tumor cells and normal cells in vitro and the effect on the growth curve of KB and KBV200 cells were observed by MTT assay. The apoptosis of CIP-36 cells was detected by fluorescent staining with Hoechst and the effect of CIP-36 on KB and KBV200 The effect of CIP-36 on the expression of P-gp in KBV200 cells was observed by immunohistochemistry. The changes of p53, p21, caspase-3, bax, mdr-1 and bcl- The results showed that CIP-36 had a good inhibitory effect on a variety of tumor cells, and had a significant inhibitory effect on drug-resistant strains of cells. Fluorescent staining showed that CIP-36 induced apoptosis in KBV200 cells. At the same time, CIP-36 dose-dependently increased the mRNA expression of p53, p21, caspase-3 and bax in KBV200 and KB cells and decreased the expression of mdr-1 and bcl-2 mRNA significance. Immunohistochemistry results showed that CIP-36 significantly reduced P-gp expression in KBV200. It is suggested that CIP-36 may overcome the multi-drug resistance of tumor cell lines by affecting the expression of multiple genes and proteins related to drug resistance.