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五味子甲素对前列腺癌细胞增殖、凋亡及Hippo-YAP信号通路的影响

Impacts of Schisandrin A on proliferation, apoptosis and Hippo-YAP signaling pathway of prostate cancer cell

摘要目的:探讨五味子甲素对前列腺癌细胞增殖、凋亡的影响及其作用机制。方法:培养人前列腺癌DU145细胞并分为DU145组(正常培养)、五味子甲素L组(加入50 μmol/L五味子甲素)、五味子甲素M组(加入100 μmol/L五味子甲素)、五味子甲素H组(加入150 μmol/L五味子甲素)和辛伐他汀组(加入50 μmol/L辛伐他汀)。显微镜下观察各组细胞形态,CCK8法检测细胞增殖能力,细胞划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力,流式细胞术检测细胞凋亡情况,Western blotting法检测磷酸化(p)-哺乳动物STE20样蛋白激酶1(MST1)、MST1、p-大肿瘤抑制因子1(LATS1)、LATS1、p-Yes相关蛋白(YAP)及YAP蛋白表达。计量资料以均数±标准差( ± s)表示,多组间比较采用单因素方差分析,两组间比较采用 t检验。 结果:与DU145组对比,五味子甲素L、M、H组及辛伐他汀组细胞数量减少且细胞逐渐皱缩、间距变大,细胞存活率[(100.00±0.00)%、(88.41±9.36)%、(62.34±7.31)%、(42.57±5.01)%、(45.47±5.65)%]、迁移[(90.11±13.43)%、(74.16±8.08)%、(57.53±7.34)%、(41.34±6.79)%、(43.44±5.26)%]和侵袭能力[(89.01±10.31)%、(73.11±9.23)%、(55.62±7.67)%、(41.13±6.35)%、(40.36±5.68)%]及p-YAP/YAP蛋白表达水平(0.98±0.08、0.83±0.11、0.69±0.07、0.55±0.07、0.53±0.05)显著下降,凋亡率[(2.88±0.34)%、(5.20±0.57)%、(8.37±0.94)%、(12.71±1.58)%、(12.03±2.21)%]和p-MST1/MST1(0.41±0.04、0.53±0.07、0.75±0.07、0.89±0.08、0.88±0.07)、p-LATS1/LATS1蛋白表达水平(0.40±0.04、0.52±0.06、0.64±0.06、0.77±0.08、0.79±0.08)显著提高,差异均具有统计学意义( P<0.05)。 结论:五味子甲素可能通过抑制Hippo-YAP信号通路抑制前列腺癌细胞增殖,促进细胞凋亡。

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abstractsObjective:To investigate the effects of Schisandrin A on the proliferation and apoptosis of prostate cancer cell and its mechanism.Methods:Human prostate cancer DU145 cell were cultured in vitro and grouped into DU145 group (normal culture), Schisandrin A L group (50 μmol/L Schisandrin A was added), Schisandrin A M group (100 μmol/L Schisandrin A was added), Schisandrin A H group (150 μmol/L Schisandrin A was added) and Simvastatin group (50 μmol/L Simvastatin was added). Cell morphology of each group was observed under microscope, cell proliferation ability was detected by CCK8 method, cell migration ability was detected by cell scratch assay, cell invasion ability was detected by Transwell assay, and cell apoptosis was detected by flow cytometry, the expression of phosphorylation (p) - mammalian STE20-like protein kinase 1 (MST1), MST1, p-large tumor suppressor 1 (LATS1), LATS1, p-Yes associated protein (YAP) and YAP protein were detected by Western blotting. Measurement data were expressed as mean± standard deviation ( ± s), one-way ANOVA for comparisons between multiple groups, and t-test for comparisons between two groups. Results:Compared with DU145 group, the number of cells in Schisandrin A L, M, H groups and Simvastatin group decreased, and the cells gradually shrunk and the spacing became larger, the cell survival rate [(100.00±0.00)%, (88.41±9.36)%, (62.34±7.31)%, (42.57±5.01)%, (45.47±5.65)%], migration [(90.11±13.43)%, (74.16±8.08)%, (57.53±7.34)%, (41.34±6.79)%, (43.44±5.26)%] and invasion [(89.01±10.31)%, (73.11±9.23)%, (55.62±7.67)%, (41.13±6.35)%, (40.36±5.68)%], and the expression of p-YAP/YAP protein (0.98±0.08, 0.83±0.11, 0.69±0.07, 0.55±0.07, 0.53±0.05) were significantly decreased, the apoptosis rate [(2.88±0.34)%, (5.20±0.57)%, (8.37±0.94)%, (12.71±1.58)%, (12.03±2.21)%] and the expression of p-MST1/MST1 (0.41±0.04, 0.53±0.07, 0.75±0.07, 0.89±0.08, 0.88±0.07] and p-LATS1/LATS1 protein (0.40±0.04, 0.52±0.06, 0.64±0.06, 0.77±0.08, 0.79±0.08) were significantly increased, and the differences were statistically significant ( P<0.05). Conclusion:Schisandrin A may inhibit the proliferation of prostate cancer cell and promote cell apoptosis by inhibiting Hippo-YAP signaling pathway.

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