志苓胶囊对小细胞肺癌细胞株NCI-H446细胞凋亡及h-TERT、CD44表达的影响

目的研究志苓胶囊(抗癌复方Ⅱ号)对人小细胞肺癌细胞株NCI-H446诱导凋亡作用机制以及对细胞黏附分子表达的影响。方法将志苓胶囊按其不同的中西药成份配制成相应的中药组、西药组和复方组,与NCI-H446共培养后,倒置显微镜观察、Hochest33258荧光染色法检测凋亡细胞,RT-PCR、Western blot、流式细胞仪分别检测bcl-2、bax、hTERT基因mRNA和相应蛋白表达水平变化以及黏附分子CD44的表达变化。结果NCI-H446经不同的药物组处理后,倒置显微镜、Hochest33258荧光染色可观察到细胞凋亡形态学改变,bcl-2基因mRNA和蛋白表达水平均有不同程度下降,bax基因表达水平则有不同程度增强,复方组改变最明显,各药物组与对照组比较差异具有显著性(P<0.01)。中药组和西药组hTERT基因和蛋白表达水平未见明显改变,复方组表达水平下调,与对照组比较差异具有显著性(P<0.01)。各药物组CD44表达水平降低,其中复方组降低水平最明显(P<0.01)。结论志苓胶囊可能通过下调NCI-H446细胞bcl-2、hTERT基因和CD44分子表达,增强bax基因表达,从而诱导NCI-H446细胞凋亡,抑制细胞黏附、转移。

Effects of Zhiling capsule on apoptosis and expressions of hTERT, CD44 in small cell lung cancer cell line NCI-H446

Aim To explore the mechanisms of the apoptosis induction and the effects of adhesion suppression of Zhiling capsule (ZLJN) in small cell lung cancer cell line NCI-H446.Methods According to the different components of ZLJN,NCI-H446 cells were treated with traditional Chinese medicine,western medicine and ZLJN composite groups.Apoptotic cells were tested by light microscopy,Hochest33258 staining method.The mRNA and protein expressions of bcl-2,bax and hTERT were analyzed by RT-PCR and Western blot respectively.The expressions of CD44 were detected by flow cytometry.Results After NCI-H446 cells were treated with different drug groups,The morphological changes of apoptotic cells were found by light microscopy and Hochest33258 staining method.The mRNA and protein expressions of bcl-2 were down-regulated while the expressions of bax were up-regulated compared to the control groups(P<0.01).The expressions of hTERT in traditional Chinese medicine and western medicine groups show no changes,but the expression in composite group was significantly lowered compared to the control group(P<0.01).The expressions of CD44 were also decreased in each group.All the changes were the most significant in composite group(P<0.01).Conclusion ZLJN could efficiently induce apoptosis in NCI-H446 cells and suppress the cell adhesion and metastasis,which maybe related to suppression of the expressions of bcl-2,hTERT and CD44 and activation of the expression of bax.