微小RNA-335靶向特异蛋白一员对前列腺癌细胞增殖的调控作用  

microRNA -335 regulate cell proliferation by targeting specific protein one in prostatic cancer cells lymph node carcinoma of prostate

作　　者：王浦[1] 车梓[2] 

机构地区：[1]山东省菏泽市立医院泌尿外科,菏泽274000 [2]山东省淄博市中心医院泌尿外科,淄博255036

出　　处：《中华实验外科杂志》2016年第6期1494-1496,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察微小RNA（miRNA，miR）-335靶向特异蛋白-员（Sp1）对前列腺癌细胞（LNCaP）增殖的调控。方法选取LNCaP为研究对象，采用荧光素酶活性实验验证miR-335对Sp1的靶向作用；采用反转录-聚合酶链反应（RT—PCR）检测细胞中miR-335表达；采用Westernblot检测细胞中Sp1蛋白表达；采用噻唑蓝（MTr）法检测LNCaP细胞增殖活性。结果与前列腺增生上皮细胞BPH．1比较，LNCaP中miR-335表达（0．83±0．22）显著低于BPH-1（2．97±0．51）；LNCaP中Sp1蛋白表达（0．92±0．18）显著高于BPH-1（0．31±0．07）。miR-335可与Sp13’-非编码区（UTR）特异性结合，降低荧光值。转染miR-335可使LNCaP中miR-335高表达，降低Sp1蛋白表达，并抑制LNCaP细胞增殖活性。同时转染miR-335及Sp1表达质粒可增加LNCaP中Sp1蛋白表达，并部分逆转miR-335对LNCaP细胞增殖的抑制作用。结论miR-335可通过靶向Sp1抑制前列腺癌细胞LNCaP增殖。Objective To evaluate the effect of microRNA （miRNA, miR） -335 target regulating Specific protein one （ SpI ） on the cell proliferation of prostatic cancer cells lymph node carcinoma of pros- tate （LNCaP）. Methods Chose prostatic cancer cells LNCaP as research object. Luciferase activity ex- periment was used to detect the target effect of miR - 335 to Spl. Reverse transcriptase - polymerase chain reaction （ RT - PCR） was used to detect the mRNA expression of miR - 335. Western blotting was used to detect the protein expression of Sp1. Methyl thiazol tetrazolium （MTF） method was used to detect the cell proliferation activity of LNCaP. Results Compared with prostate hyperplasia epithelial cell BPH - 1, the miR -335 expression level of LNCaP （0. 83 ±0. 22） is significantly lower than BPH - 1 （2. 97 ±0. 51 ）, and the Spl protein expression of LNCaP （0.92 ± 0. 18 ） is significantly higher than BPH - 1 （0. 31 ± 0.07）. miR- 335 specific combined with Spl 3＇ -untranslated region （UTR）, and reduced fluorescence value. After transfection miR- 335 to LNCaP, the miR -335 expression significantly increased, the Spl protein expression significantly reduced, and cell proliferation was significantly restrained. After transfec- tion miR- 335 and Spl to LNCaP, the Spl expression significantly increased, and partly reversed the in- hibitory effect of miR - 335 on cell proliferation. Conclusion miR - 335 can target Spl to inhibit the pro- liferation activity of prostatic cancer cells LNCaP.

关 键 词：微小RNA-335 特异蛋白一员 前列腺癌 靶向 细胞增殖 

分 类 号：R737.25[医药卫生—肿瘤]