PTEN基因过表达对前列腺癌PC-3细胞增殖的影响

Effect of PTEN Gene Overexpression on the Proliferation of PC-3 Cells in Patients with Prostate Cancer

目的构建PTEN基因的表达载体并转染前列腺癌PC-3细胞,探讨其对前列腺癌PC-3细胞增殖的调节作用。方法设计合成针对PTEN PCR引物序列,构建含有PTEN基因的过表达载体。应用Real Time PCR及Western blot检测重组质粒刺激PC-3细胞后PTEN的mRNA和蛋白的表达水平,应用MTT法检测过表达载体对PC-3细胞增殖的调节作用。结果经酶切、GFP表达证实,成功构建pSicoR-PTEN载体;Real Time PCR及Western blot结果显示pSicoR-PTEN载体可在PC-3细胞内成功表达PTEN,并且mRNA和蛋白的表达量相对于空质粒组和PC-3对照组显著上升(P<0.05);MTT结果显示PTEN的过表达对PC-3细胞增殖有明显的抑制作用(P<0.05)。结论成功构建的PTEN过表达载体可以有效的提高PC-3细胞内PTEN的mRNA和蛋白表达水平,并可抑制PC-3细胞的增殖。

cancer cell Objective To construct the recombinant lentivirus vector and to explore its effects on prostate line PC -3. Methods The PCR primers sequence targeting to PTEN gene was designed and was cloned to pSicoR vector. The recombinant lentivirus vector was used to challenge PC -3 cells. The expression level of PTEN mRNA and protein was detected by Real Time PCR and Western blotting. The effects on PC - 3 cells proliferation were analyzed by MT-F assay. Results The restriction enzyme digestion, DNA sequencing and detection of GFP expression demonstrated that recombinant lentiviru., vector was constructed successfully. Real Time PCR and Western blot analysis confirmed that the expression of PTEN mRNA and protein were ele- vated. The MTI＂ assay showed that the anti - proliferation effect of lentivirus vector of PTEN on PC - 3 cells were significant（ P 〈 0.05 ）. Conclusions The recombinant lentivirus vector could effectively elevated the mR- NA and protein expression of PTEN in PC -3 cells and depress the proliferation of PC -3 cells.