本文選題：有絲分裂驅(qū)動(dòng)蛋白 + 多西紫杉醇耐藥�。� 參考：《山東大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年09期

【摘要】：目的探討甲氧基-(S)-三苯甲基-L-半胱氨酸[S(MeO)TLC]靶向抑制有絲分裂驅(qū)動(dòng)蛋白(KSP)在體外試驗(yàn)中對(duì)多西紫杉醇耐藥前列腺癌的治療效果。方法培養(yǎng)建立多西紫杉醇耐藥前列腺癌細(xì)胞株(PC3R),Western blotting檢測(cè)不同細(xì)胞株(PC3、DU145及PC3R)KSP蛋白表達(dá)水平;實(shí)驗(yàn)分空白對(duì)照組、PC3R組及PC3組,MTT及臺(tái)盼蘭染色細(xì)胞活性實(shí)驗(yàn)觀察S(MeO)TLC抑制耐藥細(xì)胞增殖效果;以耐藥細(xì)胞株P(guān)C3R為研究對(duì)象,實(shí)驗(yàn)分為S(MeO)TLC組及對(duì)照組,Heochst染色、流式細(xì)胞儀及RT-PCR技術(shù)檢測(cè)S(MeO)TLC誘導(dǎo)耐藥細(xì)胞凋亡的效果。結(jié)果PC3、DU145、PC3R細(xì)胞KSP蛋白表達(dá)量差異無統(tǒng)計(jì)學(xué)意義(P0.05)。PC3R細(xì)胞S(MeO)TLC半數(shù)抑制濃度(IC50)為120 nmol/L,與PC3細(xì)胞(IC50:106 nmol/L)比較,差異無統(tǒng)計(jì)學(xué)意義(P0.05)。PC3R細(xì)胞給予S(MeO)TLC作用24 h后,87.9%細(xì)胞停滯在有絲分裂期;給藥72 h后,有絲分裂期停滯細(xì)胞顯著凋亡。與對(duì)照組相比,S(MeO)TLC組PC3R細(xì)胞Caspase-3(t=13.445,P=0.000 2)、Caspase-8(t=9.494,P=0.000 7)、Caspase-9(t=5.198,P=0.007)、PARP(t=19.097,P=0.000 04)及自噬標(biāo)志指標(biāo)LC3(t=22.609,P=0.000 02)和Beclin1(t=61.266,P=0.000 000 4)的mRNA量均明顯升高。結(jié)論前列腺癌多西紫杉醇耐藥與KSP蛋白表達(dá)無關(guān),KSP蛋白靶向抑制劑S(MeO)TLC能夠有效抑制多西紫杉醇耐藥前列腺癌細(xì)胞增殖并誘導(dǎo)其凋亡。在此過程中,內(nèi)源性和外源性Caspase依賴性的凋亡途徑均發(fā)揮了重要作用,且自噬可能發(fā)揮協(xié)同作用。
[Abstract]:Objective to investigate the therapeutic effect of methoxy-triphenylmethyl-L-cysteine [S(MeO)TLC] targeting mitogen inhibitor KSPin on docetaxel resistant prostate cancer in vitro.Methods the expression levels of PC3DU145 and PC3R)KSP protein were detected by Western blotting in different cell lines of docetaxel resistant prostate cancer cell line (PC3RX).The effect of S(MeO)TLC on the proliferation of drug-resistant cells was observed by MTT assay and Trypan blue staining assay in the blank control group and PC3 group, and the drug resistant cell line PC3R was selected as the study object, and divided into S(MeO)TLC group and control group by Heochst staining.Flow cytometry and RT-PCR techniques were used to detect the apoptosis of drug resistant cells induced by S(MeO)TLC.Results there was no significant difference in the expression of KSP protein between PC3R cells and PC3 cells (P0.05U .PC3R cells IC50). Compared with PC3 cells (IC50: 106nmol / L), there was no significant difference in the expression of KSP protein in PC3R cells treated with S(MeO)TLC for 24 h. 87.9% of the cells stagnated in mitotic phase after being treated with S(MeO)TLC for 24 hours.72 hours after administration, the mitotic arrest cells were significantly apoptotic.Conclusion docetaxel resistance is not related to the expression of KSP protein. S(MeO)TLC can effectively inhibit the proliferation and induce apoptosis of docetaxel resistant prostate cancer cells.In this process, endogenous and exogenous Caspase dependent apoptotic pathways play an important role, and autophagy may play a synergistic role.
【作者單位】： 山東大學(xué)附屬省立醫(yī)院泌尿外科;山東大學(xué)齊魯醫(yī)院泌尿外科;
【基金】：國家自然科學(xué)基金(81202017) 山東省重點(diǎn)研發(fā)計(jì)劃(2016GSF201147) 濟(jì)南市科技發(fā)展計(jì)劃(201121060)
【分類號(hào)】：R737.25
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本文編號(hào)：1764843