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目的探讨丹参酮Ⅱa对人绒毛膜癌甲氨蝶呤耐药细胞系(JAR/MTX)的杀伤与逆转耐药效应及与甲氨蝶呤(MTX)的协同增效作用。方法根据对JAR/MTX的不同处理分为空白组、DMSO溶媒组、MTX组和丹参酮Ⅱa组。采用HE染色进行显微镜下各组细胞形态学观察;电化学发光法测定药物作用前后各组细胞β-人绒毛膜促性腺激素(β-hCG)水平;免疫组化染色法测定各组细胞热休克蛋白27(HSP27)及谷胱甘肽转移酶(GST-π)表达;MTT法检测丹参酮Ⅱa作用前后半数抑制浓度并计算其逆转耐药倍数。结果丹参酮Ⅱa组药物作用24h可见细胞核肿胀,核膜皱缩及消失,核分裂象减少及坏死,而MTX组细胞坏死不明显。GST-π主要在细胞质中表达,HSP主要在细胞核中表达。丹参酮Ⅱa组药物作用24h后HSP27的表达率为20.20%、GST-π的表达率为16.30%,与空白组比较,差异均有统计学意义(P<0.05);丹参酮Ⅱa组药物作用24h后β-hCG为(23.04±11.32)U/L,与空白组[(358.80±19.00)U/L]比较,差异有统计学意义(P<0.05);丹参酮Ⅱa与MTX配伍作用于JAR/MTX细胞后的逆转耐药倍数为5.3。结论丹参酮Ⅱa对人绒毛膜癌耐药细胞株JAR/MTX具有杀伤与逆转耐药的效应,丹参酮Ⅱa可使JAR/MTX细胞HSP27及GST-π的表达明显下调,并且与MTX同时应用具有协同增效作用。
Objective To investigate the killing effect of Tanshinone Ⅱa on human choriocarcinoma cell line MTX (MTAR) and the reversal of drug resistance and synergism with methotrexate (MTX). Methods According to the different treatment of JAR / MTX, the rats were divided into blank group, DMSO vehicle group, MTX group and tanshinone Ⅱ a group. The morphological changes of cells in each group were observed by HE staining. The level of β-human chorionic gonadotropin (β-hCG) in each group was determined by electrochemiluminescence. The heat shock Protein 27 (HSP27) and glutathione transferase (GST-π) were detected by MTT assay. The half inhibitory concentration (TCR) Results The tanshinone Ⅱa group showed swelling of the nucleus, shrinkage and disappearance of the nuclear membrane, reduction and necrosis of mitosis, and no obvious cell necrosis in MTX group. GST-π is mainly expressed in the cytoplasm, and HSP is mainly expressed in the nucleus. Tanshinone Ⅱ a group 24h after the HSP27 expression rate of 20.20%, GST-π expression rate of 16.30%, compared with the blank group, the difference was statistically significant (P <0.05); tanshinone Ⅱ a group of drugs 24h after β (23.04 ± 11.32) U / L, respectively. Compared with the blank group [(358.80 ± 19.00) U / L], the difference was statistically significant (P <0.05); when tanshinoneⅡa and MTX were used in JAR / MTX cells The reversal of resistance was 5.3. Conclusion Tanshinone Ⅱa can kill or reverse drug-resistant JAR / MTX cell line in human choriocarcinoma cell line. Tanshinone Ⅱa can down-regulate the expression of HSP27 and GST-π in JAR / MTX cell line, and in combination with MTX, Effective effect.