摘要
目的 观察11-19赖氨酸富集白血病基因2(ELL2) 在人前列腺癌组织中表达水平及其对人前列腺癌细胞株C4-2增殖的影响.方法 免疫组织化学方法观察ELL2在人前列腺癌组织中的表达,探讨ELL2与前列腺癌恶性程度的关系;用人工合成雄激素R1881刺激人前列腺癌细胞株C4-2,在不同时间点检测其对雄激素的反应性;分别对C4-2细胞株转染ELL2特异性干扰RNA(ELL2-siRNA)及过表达ELL2质粒,噻唑蓝(MTT)实验评估干预细胞ELL2表达后细胞增殖能力的变化,溴脱氧尿嘧啶核苷(BrdU)掺入实验评估细胞DNA合成能力的变化.结果 ELL2在人前列腺癌组织中表达水平明显降低,且随着前列腺癌恶性程度的增加进一步降低;对ELL2及前列腺癌的Gleason评分进行相关分析显示,两者呈负相关(r=0.836,P=0.000).R1881可在48h内促进ELL2的蛋白表达,在24h及48h的蛋白表达量分别增加(38.27±10.11)%(t=7.573,P=0.005)、(58.34±13.70)%(t=8.519,P=0.003).C4-2细胞转染ELL2特异性干扰RNA后,ELL2在蛋白水平明显降低,两条干扰RNA组的细胞ELL2蛋白表达分别下降(56.38±11.26)%(t=10.01,P=0.002)、(65.27±12.10)%(t=10.790,P=0.002);转染过表达ELL2质粒后,ELL2在蛋白水平明显升高,过表达组的细胞ELL2蛋白增加(80.23±20.94)%(t=7.662,P=0.005).转染ELL2特异性干扰RNA使C4-2细胞增殖能力明显升高,两条干扰RNA组的细胞增殖能力分别升高(39.28±8.52)%(t=11.300,P=0.000)、(48.20±7.87)%(t=15.000,P=0.000);转染过表达ELL2质粒使细胞增殖能力降低,过表达组的细胞增殖能力降低(42.74±10.38)%(t=10.090,P=0.000).转染ELL2特异性干扰RNA使细胞DNA合成能力增加,而转染过表达ELL2组的DNA合成能力降低.结论 ELL2在人前列腺癌组织中低表达,且与其恶性程度呈负相关,ELL2可在一定程度上抑制前列腺癌细胞的增殖能力.
Objective To observe the level of eleven-nineteen lysine-rich leukemia gene 2 (ELL2) in human prostate cancer tissues and its effect on proliferation of prostate cancer cell C4-2.Methods The expression of ELL2 in human prostate cancer tissues was analyzed by immunohistochemical method, and the relationship between ELL2 and malignancy degree of prostate cancer was evaluated.The human prostate cancer cell line C4-2 was stimulated with synthetic androgen R1881 to detect its response to androgen at different time points.ELL2-small interfering RNA (siRNA) and overexpression of ELL2-bearing plasmid were transfected into C4-2 cells respectively, then cell proliferation was evaluated by methyl thiazol tetrazolium (MTT) assay.The DNA synthesis ability of cells was assessed by 5-Bromo-2'-deoxy-uridine (BrdU) incorporation assay.Results The expression of ELL2 was significantly lower in human prostate cancer than that of benign specimen, and decreased with the increase of malignancy of prostate cancer.The correlation analysis between ELL2 and prostate cancer Gleason score reveal a negative correlation result (r=0.836, P=0.000).The protein expression of ELL2 was significantly increased in 48 h and increased by (38.27±10.11)% (t=7.573, P=0.005) and (58.34±13.70)% (t=8.519, P=0.003) at 24 h and 48 h, respectively.The expression of ELL2 protein in the two RNA interference groups were significantly decreased, and the decreased rate were (56.38±11.26)% (t=10.010, P=0.002) and (65.27±12.10)% (t=10.790, P=0.002), respectively.While the ELL2 protein expression in the overexpression group was significantly increased at (80.23±20.94)% (t=7.662, P=0.005).The proliferation ability of C4-2 cells was significantly increased (39.28±8.52)% (t=11.300, P=0.000) and (48.2±7.87)% (t=15.000, P=0.000) in the two RNA interference groups, respectively, compared with the control group.While its proliferation ability decreased (42.74±10.38)% (t=10.090, P=0.000) in ELL2-bearing plasmid.Transfection of ELL2-specific interfering RNA increased the ability of DNA synthesis, while the transfection of ELL2-bearing plasmid cells had lower DNA synthesis ability.Conclusion ELL2 is lowly expressed in human prostate cancer tissues and negatively correlated with the malignant degree.ELL2 can inhibit the proliferation of prostate cancer cells to a certain extent.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2017年第3期433-436,共4页
Chinese Journal of Experimental Surgery
基金
河南省科技攻关计划资助项目(162102310040)