丹参酮ⅡA对人胃癌MKN-45细胞增殖及基质金属蛋白酶-2表达的影响

Effects of TanshinoneⅡA on Proliferation and MMP-2 mRNA Expression in Human Gastric Carcinoma Cell Line MKN-45

[目的]通过观察丹参酮ⅡA对人胃癌MKN-45细胞的抑制作用及对转移相关基因MMP-2表达的影响,探讨其抗肿瘤的机制。[方法]MTT法检测不同浓度丹参酮ⅡA作用下,体外培养的MKN-45细胞的增殖情况;RT-PCR法、酶谱分析法检测MKN-45细胞MMP-2 mRNA及蛋白的表达。[结果]丹参酮ⅡA各浓度组对肿瘤细胞增殖的抑制率与阴性对照组相比,有显著差异(P<0.05),相关回归分析表明24h、48h、72h的半数抑制浓度分别为42.5μg/ml、19.4μg/ml和7.4μg/ml。PCR半定量分析发现:丹参酮ⅡA各组的MMP-2 mRNA表达均受到抑制,抑制程度随着药物浓度的增加而增加,酶谱分析法显示:丹参酮ⅡA能降低胃癌MKN-45细胞的MMP-2蛋白表达,其作用呈剂量-效应正相关,与阴性对照组相比,差异有显著性(P<0.05)。[结论]丹参酮ⅡA能有效抑制胃癌MKN-45细胞的生长,并具有明显的时间和剂量依赖性。丹参酮ⅡA可能通过抑制胃癌MKN-45细胞MMP-2 mRNA的转录下调MMP-2蛋白的表达,进而抑制肿瘤增殖。

[Objeetive] To determine the anti-tumor mechanism of Tanshinone ⅡA on human Gastric Carcinoma Cell Line MKN- 45. [Methods]MKN-45 cells were routinely cultured in vitro. The MTT assay had been developed for quantitative evaluation of the proliferation of MKN-45 cells of each group. Expression of MMP 2 mRNA was measured by RT-PCR and Zymography. [Results] The difference of inhibition rates（IR） on Cell Line MKN-45 between the negative and Tan ⅡA group was significant（P〈0. 05）. The expression of MMP-2 mRNA of Tanshinone ⅡA groups（3,6,9,12μg/ml） was lower than that of the negative group, statistical difference between the groups above was significant（P（0.05）, except for the 3μg/ml one. [Conclusions] Tanshinone ⅡA inhibits the growth of Cell Line MKN-45 in a dose and time-dependent manner, which may be realized through downregulating the expression of MMP-2.