熊果酸诱导子宫内膜癌细胞HHUA凋亡的研究

Ursolic acid induces apoptosis in endometrial carcinoma HHUA cells

目的探讨三萜类化合物熊果酸(ursolic acid,UA)诱导子宫内膜癌细胞HHUA的凋亡作用。方法采用细胞培养技术,用不同浓度的药物在一定的时间内处理细胞株,采用MTT法、活细胞荧光染色技术、流式细胞技术(FCM)、RT-PCR、细胞免疫化学技术,研究UA对HHUA细胞的增殖抑制和诱导凋亡作用。结果UA抑制HHUA细胞增殖,且呈时间和浓度依赖性,并使细胞呈典型的凋亡形态学特征,核质浓集,出现凋亡小体,使细胞周期阻滞在G0/G1期,并诱导人半胱天冬蛋白酶3(caspase-3)蛋白表达增加,bcl-2蛋白表达减少。结论UA诱导HHUA细胞凋亡,其机制涉及caspase-3和bcl-2依赖性凋亡调节通路。

Objective To study the effects of ursolic acid （UA） on the growth of the endometrial cancer cell line HHUA. Methods The HHUA cells were treated with ursolic acid at different concentrations. The proliferation inhibition was determined by MTF assay. Morphological examination and flow cytometry were used to determine the apoptosis. RT-PCR and the immunohistochemical method were used to determine the expressions of apoptosis related genes caspase- 3 and bcl-2. The semi-quantification of protein expression was analyzed by pathological image-analysis. Results UA strongly inhibited the proliferation of HHUA cells. Apoptosis was induced by UA treatment. The morphology of HHUA showed changes such as chromatin aggregation, nuclear and cytoplasmic condensation and apoptotic body appearance. UA caused significant G0/G1 phase arrest with a concomitant decrease of cell population in S and GriM phases. Apoptotic cells were found by flow cytometry. The maximal early apoptotic rate was （29.1 ± 2.43） % when exposed to 40 umol/L for 72 h. The expression of the caspase-3 gene was enhanced while the expression of the anti-apoptotic gene bcl-2 was decreased. All these effects were in a dose- and time-dependent manner. Conduslon UA induces the apoptotic processes in the endometrial cancer HHUA cell line by involving the caspase-3 pathway and bcl-2 family proteins.