Numb基因调控前列腺癌PC-3细胞生长的研究

Growth of prostate cancer PC-3 cell regulated by Numb

目的探讨Numb基因与前列腺癌发生发展的关联性及其与p53、Notch信号通路之间的关系。方法通过构建Numb过表达质粒,转染人PC-3细胞(CRPC细胞),建立稳转细胞株并验证,通过CCK-8检测肿瘤细胞的增殖水平;通过免疫印迹检测过表达Numb后对细胞中p53与Notch1的表达影响;将PC-3细胞植入裸鼠皮下,建立人去势抵抗性前列腺癌裸鼠移植瘤模型;计算并绘制肿瘤的生长曲线,通过流式细胞技术检测细胞的生长周期,并对移植瘤进行病理鉴定。结果过表达Numb可抑制PC-3细胞增殖水平。过表达Numb可抑制PC-3细胞中Notch1表达、促进p53的表达。过表达Numb可抑制裸鼠种植瘤的生长,主要是通过减少PC-3细胞中S期细胞比例,并且免疫组化也证实过表达Numb可抑制肿瘤中Notch1表达并促进p53表达。结论Numb基因的过表达会促进抑癌基因p53的激活以及癌基因Notch表达下调,抑制前列腺癌的发生发展。

Objective To investigate the relationship between Numb and the development and progression of prostate cancer and its relationship with P53 and Notch signaling pathways.Methods Numb overexpressed plasmid was constructed and transfected into human PC-3 cells(CRPC cells)to establish stable cell lines.The proliferation level of cell line was detected by CCK-8.The expression of p53 and Notchl in Numb-expressed cells after overexpression was detected by Western blot.PC-3 cells were implanted subcutaneously into nude mice to establish the xenograft model of castration-resistant prostate cancer in nude mice.The tumor growth curve was calculated and plotted,the cell growth cycle was detected by flow cytometry,and the transplanted tumor was pathologically identified.Results Overexpression of Numb can inhibit proliferation leve of PC-3 cells;Overexpression of Numb inhibits Notchl and promotes P53 expression in PC-3 cells;Numb overexpression inhibits the growth of Numb-expressed PC-3 cells in nude mice by decreasing the proportion of S phase cells in PC-3 cells.Overexpression of Numb also inhibits Notchl and promotes P53 expression in nude mice.Conclusion Numb gene overexpression can promote the activation of cancer suppressor gene p53 and decrease the expression of oncogene Notch,and inhibit the progression of prostate cancer.