CAJ | 학술논문

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.
목적 관찰제10호염색체결실적린산매급장력단백동원물기인(PTEN)대인전렬선암세포주PC-3전이급기질금속단백매(MMP)-2、MMP-9표체적영향.방법 이휴대야생형PTEN기인적선병독감염체외배양적PC-3세포,채용간접면역형광실험화Western blot실험검측PTEN、MMP-2 、MMP-9단백표체적변화;명효매보실험검측MMP-2화MMP-9매활성적변화.화흔실험화Transwell실험검측PTEN대PC-3세포체외천이화침습능력적영향.결과 Ad-PTEN감염후PC-3세포중PTEN표체명현증가,이MMP-2화MMP-9표체수평급활성균강저.화흔실험현시Ad-PTEN조세포천이거리재12h위(112.36±18.38) μm,명현소우Ad-LacZ조적(166.52±19.28) μm(t=4.55,P ＜0.01);재24 h위(214.75±35.62) μm,명현소우Ad-LacZ조적(361.87 ±46.15) μm(t=5.64,P ＜0.01);재48 h위(436.61 ±52.69) μm,명현소우Ad-LacZ조적(732.56±78.30) μm,(t=7.01,P＜0.01).Transwell실험결과현시Ad-PTEN조천입하실면적세포수[(22.40±2.41)개]명현소우Control조[(38.20±4.44)개,t=6.99,P＜0.01]급Ad-LacZ조[(37.20±4.97)개,t=5.98,P＜0.01].결론 PTEN대인전렬선암PC-3세포주전이급MMP-2、MMP-9표체유명현억제작용,제시PTEN가능재전렬선암적전이중발휘중요작용.
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.