Cyr61在前列腺癌组织中的表达及其对前列腺癌细胞增殖、上皮间质转

广东医学 2019 年6 月 第 40 卷第 11 期 Guangdong Medical Journal Jun. 2019, Vol. 40, No. 11?1535?Cyi61在前列腺癌组织中的表达及其对前列腺癌

细胞增殖、上皮间质转化和自噬的影响高国栋-张庆云S易利駕李岩2天津市蓟州区人民医院'预防保健部，2浹尿外科(天津301900)【摘要】目的探讨富含半胱氨酸蛋白61(Cyi61)在前列腺癌组织中的表达及其对前列腺癌细胞增殖、 上皮间质转化和自噬的影响。方法收集手术切除的前列腺癌组织和癌旁正常组织，分别使用免疫组织化

学染色、荧光定量聚合酶链式反应(RT-PCR)和蛋白质免疫印迹(WB)检测组织中CyWl的表达。将PC -3 细胞分为空白组、siControl组和siCyr61组,siCyi61组使用RNA干扰将Cyi61沉默,siControl组和空白组分别

使用无关序列和PBS作为对照。qRT - PCR检测细胞中Cyi61 mRNA的表达,WB检测细胞中Cyi61、E -钙黏 蛋白、波形蛋白、p-PI3K、p-AKT蛋白的表达，免疫荧光染色检测细胞中LC - 3 II的表达，MTT试验检测细 胞增殖情况。结果 CyWl在前列腺癌旁组织中呈阴性或低表达，而在前列腺癌组织中的表达水平升高。与

癌旁组织相比较，前列腺癌组织中Cyi61 mRNA和蛋白质的表达水平升高，差异有统计学意义(P < 0. 05 )。

与空白组和siControl组相比较,siCyr61组细胞中Cyi61 mRNA和蛋白质明显降低，差异有统计学意义(P< 0. 05)；在培养24、48、72 h时0D值降低(P<0.05) ；E -钙黏蛋白的表达升高，波形蛋白的表达降低(P < 0.05)；LC-3U的荧光强度较高:p-PI3K和p - AKT的表达降低(P<0.05)o结论 Cyi61在前列腺癌患者

的癌组织中高表达，使用RNA干扰将Cyi61沉默后可以抑制人前列腺癌PC-3细胞的增殖，降低上皮间质转

化相关蛋白的表达，促进自噬的发生，可能与对PI3K/AKT信号通路的抑制有关。【关键词】 富含半胱氨酸蛋白61；前列腺癌；RNA干扰；上皮间质转化；自噬【中图分类号】R737.25；Q2

【文献标志码】ADOI：10. 13820/j. cnki. gdyx. 20190117Expression of Cysteine - rich Protein 61 in Prostate Cancer and Its Effect on Proliferation, Epithelial Mesenchy-

mal Transition and Autophagy of Prostate Cancer Cells. GAO Guo - dong\ , ZHANG Qing -yun, YI Li, LI Yan.Department of Preventive Heath, Tianjin Jizhou People's Hospital, Tianjin 301900, China[Abstract ] Objective To explore the expression of cysteine 一 rich protein 61 ( Cyr61) in prostate cancer and its

effect on proliferation, epithelial mesenchymal transition ( EMT) and autophagy of prostate cancer cells. Methods Im-

munohistochemical staining, quantitative real time polymerase chain reaction ( RT - PCR) and western blot ( WB) were used to detect the expression of Cyi61 in resected prostate cancer tissues and adjacent normal tissues. PC - 3 cells were

divided into blank group, siControl group and siCyr61 group. Cyi61 gene was silenced by RNA interference in siCyi61

group, and unrelated sequences and PBS were used as controls in siControl group and blank group, respectively. qRT -

PCR was used to detect the expression of Cyi61. WB was used to detect the expression of Cyi61, E 一 cadherin, vimentin, p - PI3K and p - AKT. Immunofluorescence staining was used to detect the expression of LC — 3II. MTT was used to ex-

amine cell proliferation. Results Cyr61 expression was negative or low in the adjacent tissues of prostate cancer, but in-creased in prostate cancer. Compared with the adjacent tissues, the expression of Cyi61 in prostate cancer tissues was sig-

nificantly increased (P <0. 05). Compared with the blank group and siControl group, in siCyi61 group the expression of Cyi61 decreased； OD decreased at 24 h, 48 h and 72 h； the expression of E - cadherin increased； the expression of vim-

entin decreased ； LC - 311 fluorescence intensity was higher ； and the expression of p - PI3K and p - AKT decreased ( P < 0. 05). Conclusion Cyi61 is highly expressed in cancer tissues of prostate cancer patients. Silencing Cyi61 with RNA

interference can inhibit the proliferation of human prostate cancer PC - 3 cells, reduce the expression of EMT - related proteins and promote autophagy, which may be related to the inhibition of PI3K/AKT signaling pathway.[Key words ] Cysteine - rich angiogenic inducer 61 ( CyWl ) ； Prostatic cancer； RNA interference; Epithelial

mesenchymal transition ； Autophagy前列腺癌是主要发生于中年和老年男性的恶性 有恶性肿瘤的第2位⑷。富含半胱氨酸蛋白61肿瘤，在全世界的发病率为25. 3/10万，位于男性所

(cysteine - rich angiogenic inducer 61 ,Cyr61)是一种