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目的:使用小干扰RNA(siRNA)抑制人前列腺癌LNCa P细胞中婆罗双树样基因4(SALL4)的表达并对抑制效果进行测定,检测沉默(SALL4)后LNCa P细胞增殖、集落形成及凋亡等生物学行为的变化。方法:培养LNCa P细胞,分为si-SALL4组、阴性对照组与空白对照组。Real-time PCR与Western blotting技术检测SALL4 mRNA与蛋白水平。采用MTS比色法观察各组细胞增殖能力的变化,采用集落形成实验观察各组细胞集落形成能力的变化,流式细胞术研究SALL4对细胞凋亡的影响,利用Western blotting方法检测凋亡相关蛋白Bax和Bcl-2的表达情况。结果:与阴性对照组相比,si-SALL4组SALL4 mRNA和蛋白表达水平、细胞增殖能力、细胞集落形成能力均明显降低(P<0.05),而凋亡细胞明显增加(P<0.05),Bcl-2的表达减弱,而Bax的表达增强。结论:通过siRNA技术沉默(SALL4)表达可抑制前列腺癌LNCa P细胞增殖和集落形成能力,促进细胞凋亡,其促进凋亡作用可能与Bcl-2及Bax表达有关。
OBJECTIVE: To inhibit the expression of SALL4 in human prostate cancer LNCa P cells by using small interfering RNA (siRNA) and determine the inhibitory effect of SALL4 on the proliferation and colony formation of LNCa P cells after SALL4 Death and other biological behavior changes. Methods: LNCa P cells were cultured and divided into si-SALL4 group, negative control group and blank control group. Real-time PCR and Western blotting were used to detect SALL4 mRNA and protein levels. The changes of cell proliferation were observed by MTS colorimetric assay. The colony formation ability of each group was observed by colony formation assay. The effect of SALL4 on apoptosis was analyzed by flow cytometry. The apoptosis-related protein Bax and Bcl-2 expression. Results: Compared with the negative control group, the expression of SALL4 mRNA and protein, cell proliferation and colony forming ability in si-SALL4 group were significantly decreased (P <0.05), while apoptotic cells were significantly increased (P <0.05) -2 expression decreased, while the expression of Bax increased. CONCLUSIONS: SALL4 expression can inhibit the proliferation and colony-forming ability of prostate cancer LNCa P cells and promote apoptosis. The apoptosis-promoting effect may be related to the expression of Bcl-2 and Bax.