本文選題：前列腺癌 + 新生霉素�。� 參考：《現(xiàn)代腫瘤醫(yī)學(xué)》2016年22期

【摘要】：目的:通過體外細(xì)胞實驗,觀察新生霉素是否抑制了前列腺癌PC-3細(xì)胞的增殖,并在分子水平和形態(tài)學(xué)上證實新生霉素通過細(xì)胞自噬抑制了前列腺癌PC-3細(xì)胞的增殖。為新生霉素應(yīng)用于去勢抵抗性前列腺癌(castration-resistant prostate cancer,CRPC)的臨床治療提供理論和實驗依據(jù)。方法:設(shè)置對照組和不同濃度的新生霉素實驗組,作用前列腺癌PC-3細(xì)胞24小時后,利用MTT法測定對照組和不同濃度實驗組的增殖率,并計算出新生霉素的半抑制率。透射電鏡觀察對照組及實驗組細(xì)胞內(nèi)的自噬體。利用免疫熒光技術(shù),在熒光顯微鏡下觀察對照組和不同濃度實驗組發(fā)生細(xì)胞自噬時的自噬標(biāo)志性蛋白LC-3Ⅱ和自噬底物蛋白p62的不同。利用RT-PCR在mRNA水平上檢測對照組和不同濃度實驗組的自噬相關(guān)基因LC-3Ⅱ、Beclin-1的表達。結(jié)果:MTT結(jié)果示新生霉素能使PC-3細(xì)胞的增殖明顯受到抑制(F=569.26,P=0.000),其抑制率隨著藥物濃度的增加而增加,根據(jù)公式計算出其IC50=1.52mmol/L。透射電鏡觀察可見實驗組PC-3細(xì)胞質(zhì)內(nèi)自噬體和自噬溶酶體數(shù)量較對照組多,且隨著濃度的增加自噬體數(shù)量增加。自噬體在細(xì)胞中的分布通過綠色熒光FITC標(biāo)記的自噬特異性蛋白LC-3Ⅱ在熒光顯微鏡下清晰顯示,在細(xì)胞膜附近有綠色熒光呈散點狀分布,隨著實驗組新生霉素濃度的增加熒光強度增加。而綠色熒光FITC標(biāo)記的自噬底物蛋白p62在熒光顯微鏡下的強度隨著藥物濃度的增加而降低。RT-PCR在mRNA水平上檢測到隨著藥物濃度的增加自噬相關(guān)基因LC-3Ⅱ、Beclin-1的表達量升高。結(jié)論:在體外細(xì)胞實驗中,新生霉素能夠在一定濃度范圍內(nèi)呈濃度依賴性的抑制PC-3細(xì)胞增殖。新生霉素能在mRNA水平上增強Beclin-1、LC-3Ⅱ的表達。新生霉素可能是通過誘導(dǎo)細(xì)胞自噬性死亡來抑制前列癌PC-3細(xì)胞的增殖。
[Abstract]:Aim: to observe whether neomycin inhibits the proliferation of prostate cancer PC-3 cells in vitro, and to prove that neomycin inhibits the proliferation of prostate cancer PC-3 cells by autophagy at molecular level and morphologically. To provide theoretical and experimental basis for the clinical treatment of castration-resistant prostate cancer with neomycin. Methods: PC-3 cells of prostate cancer were treated with different concentrations of neomycin and control group for 24 hours. The proliferation rate of control group and experimental group with different concentrations were measured by MTT assay, and the semi-inhibitory rate of neomycin was calculated. The autophagy in control group and experimental group was observed by transmission electron microscope. Immunofluorescence technique was used to observe the difference of autophagy iconic protein LC-3 鈪，

本文編號：1877921