摘要
目的 观察第10号染色体缺失的磷酸酶及张力蛋白同源物基因（PTEN）对人前列腺癌细胞株PC-3转移及基质金属蛋白酶（MMP）-2、MMP-9表达的影响.方法 以携带野生型PTEN基因的腺病毒感染体外培养的PC-3细胞,采用间接免疫荧光实验和Western blot实验检测PTEN、MMP-2 、MMP-9蛋白表达的变化;明胶酶谱实验检测MMP-2和MMP-9酶活性的变化.划痕实验和Transwell实验检测PTEN对PC-3细胞体外迁移和侵袭能力的影响.结果 Ad-PTEN感染后PC-3细胞中PTEN表达明显增加,而MMP-2和MMP-9表达水平及活性均降低.划痕实验显示Ad-PTEN组细胞迁移距离在12h为（112.36±18.38） μm,明显少于Ad-LacZ组的（166.52±19.28） μm（t=4.55,P ＜0.01）;在24 h为（214.75±35.62） μm,明显少于Ad-LacZ组的（361.87 ±46.15） μm（t=5.64,P ＜0.01）;在48 h为（436.61 ±52.69） μm,明显少于Ad-LacZ组的（732.56±78.30） μm,（t=7.01,P＜0.01）.Transwell实验结果显示Ad-PTEN组穿入下室面的细胞数[（22.40±2.41）个]明显少于Control组[（38.20±4.44）个,t=6.99,P＜0.01]及Ad-LacZ组[（37.20±4.97）个,t=5.98,P＜0.01].结论 PTEN对人前列腺癌PC-3细胞株转移及MMP-2、MMP-9表达有明显抑制作用,提示PTEN可能在前列腺癌的转移中发挥重要作用.
Objective To investigate the effects of phosphatase and tensin homolog deleted on chromosome ten （PTEN） gene expression on the prostate cancer PC-3 cells＇ metastasis and the expression of matrix metalloproteinase （MMP）-2 and MMP-9.Methods Recombinant adenovirus vectors were infected into prostate cancer cell line PC-3.The protein expression levels of PTEN,MMP-2 and MMP-9 in the three groups were detected by Western blotting and indirect immunofluorescence assay respectively.The enzymatic activities of MMP-2 and MMP-9 in the three groups were determined by gelatin zymography.Wound scratch assay was used to determine the effect of PTEN on migratory ability of PC-3 cells.Invasion ability of PC-3 cells was measured by Transwell chamber assay.Results After infection by adenovirus,the protein expression of PTEN was significantly up-regulated in PC-3 cells,and the expression and enzymatic activities of MMP-2 and MMP-9 were significantly down-regulated.Wound scratch assay showed that migration distance in Ad-PTEN group at 12,24 and 48 h was （112.36 ± 18.38）,（214.75 ± 35.62） and （436.61 ±52.69） μm respectively,which was significantly shorter than in Ad-LacZ group [（166.52 ± 19.28） μm,t=4.55,P＜0.01 at 12 h; （361.87 ±46.15） μm,t=5.64,P＜0.01 at 24 h,and （732.56 ±78.30） μm,t =7.01,P ＜0.01 at 48 h correspondingly].Transwell showed that the number of PC-3 cells that invaded the lower chamber in the Ad-PTEN group （22.40 ± 2.41） was significantly decreased as compared with the control group （38.20 ± 4.44,t =6.99,P ＜ 0.01） and the Ad-LacZ group （37.20 4.97,t =5.98,P ＜ 0.01）.Conclusion PTEN gene can significantly inhibit metastasis of the PC-3 cells and expression of MMP-2 and MMP-9,suggesting it may paly an important role in metastasis of prostate cancer.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第12期2490-2492,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81272812）
