【摘要】：目的:探討辛伐他汀對(duì)前列腺上皮細(xì)胞RWPE-1增殖及凋亡的影響。方法:設(shè)定不同濃度的辛伐他汀(0、10、20、40μmol/L)分別作用于體外培養(yǎng)的RWPE-1細(xì)胞,利用MTT法檢測(cè)細(xì)胞增殖情況,流式細(xì)胞術(shù)檢測(cè)細(xì)胞的凋亡情況。熒光定量RT-PCR檢測(cè)RWPE-1細(xì)胞的Bcl-2、Bax、Cx43 mRNA的表達(dá),Western印跡檢測(cè)Bcl-2、Bax、Cx43蛋白的表達(dá)。結(jié)果:MTT法檢測(cè)辛伐他汀(10、20、40μmol/L)作用于RWPE-1細(xì)胞72 h后,對(duì)RWPE-1細(xì)胞的抑制率分別為(21.07±6.41)%、(34.87±9.65)%和(47.18±10.88)%,與對(duì)照組[(1.21±0.54)%]比較有顯著差異(P0.05),并且呈明顯的劑量依賴關(guān)系(P0.05);處理72 h后各組的凋亡指數(shù)分別為:10μmol/L組(0.066±0.016)%,20μmol/L組(0.126±0.023)%,40μmol/L組(0.192±0.025)%,與對(duì)照組[(0.015±0.005)%]相比差異顯著(P0.01),且呈劑量依賴關(guān)系(P0.05)。熒光定量PCR檢測(cè)顯示隨著辛伐他汀濃度升高Bcl-2基因表達(dá)逐漸下調(diào)(P0.05),Bax和Cx43基因表達(dá)逐漸上調(diào)(P0.05),并且呈劑量依賴關(guān)系(P0.05)。Western印跡檢測(cè)顯示RWPE-1細(xì)胞內(nèi)Bcl-2蛋白的表達(dá)隨辛伐他汀濃度的增高逐漸下調(diào)(P0.05),Bax蛋白逐漸上調(diào)(P0.05),Cx43蛋白逐漸上調(diào)(P0.01),并且皆呈劑量依賴關(guān)系(P0.05)。Cx43的表達(dá)與Bcl-2的表達(dá)呈負(fù)相關(guān),與Bax的表達(dá)呈正相關(guān)。結(jié)論:辛伐他汀可能通過影響細(xì)胞間隙連接通訊來抑制前列腺上皮細(xì)胞增殖并誘導(dǎo)其凋亡。
[Abstract]:Aim: to investigate the effect of simvastatin on RWPE-1 proliferation and apoptosis in prostatic epithelial cells. Methods: different concentrations of simvastatin (0 10 ~ 2040 渭 mol/L) were used to treat RWPE-1 cells in vitro. Cell proliferation was detected by MTT assay and apoptosis was detected by flow cytometry. The expression of Bcl-2AXCx43 mRNA in RWPE-1 cells was detected by fluorescence quantitative RT-PCR. Western blot was used to detect the expression of Bcl-2OBaxCx43 protein. Results Simvastatin (10 ~ 20 渭 mol/L) was applied to RWPE-1 cells for 72 h. The inhibition rates of RWPE-1 cells were (21.07 鹵6.41), (34.87 鹵9.65)% and (47.18 鹵10.88), respectively, which were significantly different from those of the control group [(1.21 鹵0.54)%] (P0.05), and the apoptotic index of the control group was (0.066 鹵0.016) 渭 mol/L group (0.066 鹵0.016), (0.126 鹵0.023) 渭 mol/L group (0.192 鹵0.025) and (0.015 鹵0.005)% compared with the control group [(0.015 鹵0.005)%]. The difference was significant (P0.01), and showed a dose-dependent relationship (P0.05). Fluorescence quantitative PCR analysis showed that the expression of Bcl-2 gene decreased gradually (P0.05) and the expression of Cx43 gene increased gradually with the increase of simvastatin concentration (P0.05), and showed a dose-dependent relationship (P0.05) .Western blotting showed that the expression of Bcl-2 protein in RWPE-1 cells was increased with simvastatin. The increase of Tin-level was down-regulated (P0.05). (P0.05) the protein of Cx43 was up-regulated (P0.01), and the expression of Cx43 was negatively correlated with the expression of Bcl-2 in a dose-dependent manner (P0.05). There was a positive correlation with the expression of Bax. Conclusion: simvastatin may inhibit the proliferation and induce apoptosis of prostatic epithelial cells by affecting gap junctional communication.
【作者單位】： 福建醫(yī)科大學(xué)附屬漳州市醫(yī)院泌尿外科;
【基金】：福建省自然科學(xué)基金(2012J05164)~~
【分類號(hào)】：R697.3
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本文編號(hào)：2141345