中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2009年
2期
90-94
,共5页
刘玉兰%王燕%郎雁%吴绪峰%熊俊%朱小红%张幼鸿%张水娟%龚丽艳%卢运萍%马丁
劉玉蘭%王燕%郎雁%吳緒峰%熊俊%硃小紅%張幼鴻%張水娟%龔麗豔%盧運萍%馬丁
류옥란%왕연%랑안%오서봉%웅준%주소홍%장유홍%장수연%공려염%로운평%마정
卵巢肿瘤%反义核酸载体%增殖%侵袭%畸胎瘤细胞源性生长因子
卵巢腫瘤%反義覈痠載體%增殖%侵襲%畸胎瘤細胞源性生長因子
란소종류%반의핵산재체%증식%침습%기태류세포원성생장인자
Ovarian cancer%Antisense RNA vector%Proliferation%Invasion%PC cell derived growth factor,PCDGF
目的 观察畸胎瘤细胞源性生长因子(PCDGF)反义核酸载体对高度恶性人卵巢癌细胞株Sw626和A2780增殖和侵袭的抑制效应,并初步探讨其相关机制.方法 采用二苯基溴化四氮唑蓝(MTT)法和Boyden小窒体外侵袭实验,检测PCDGF反义RNA真核表达载体对Sw626和A2780细胞增殖和侵袭能力的影响.采用Western blot技术,检测转染PCDGF反义RNA真核表达载体前后Sw626细胞cyclin D1和CDK4蛋自表达的变化.采用逆转录聚合酶链反应(RT-PCR)和明胶酶谱法,分析PCDGF反义RNA真核表达载体对Sw626细胞基质金属蛋白酶2(MMP-2)表达和活性的影响.结果 与空白对照组相比,PCDGF反义核酸载体转染组Sw626和A2780细胞的增殖抑制率分别为72.9%和70.9%,侵袭能力分别被抑制了62.9%和59.0%.转染组Sw626细胞cyclin D1和CDK4蛋白的表达水平分别为0.38±0.08和0.37±0.13,明显低于空白对照组(0.84±0.11和0.64±0.11,P<0.01).与空白对照组(0.89±0.09)相比,转染组Sw626细胞MMP-2 mRNA的表达水平(0.66±0.11)虽未见降低(P>0.05),但MMP-2酶原的活性被叨显抑制.结论 PCDGF反义核酸可显著抑制高度恶性人卵巢癌细胞株Sw626和A2780的增殖和侵袭能力,并逆转其部分恶性表型,这可能与其能下调cyclin D1和CDK4蛋白的表达并抑制MMP-2酶原的活性有关;PCDGF可以作为卵巢癌治疗的新靶点.
目的 觀察畸胎瘤細胞源性生長因子(PCDGF)反義覈痠載體對高度噁性人卵巢癌細胞株Sw626和A2780增殖和侵襲的抑製效應,併初步探討其相關機製.方法 採用二苯基溴化四氮唑藍(MTT)法和Boyden小窒體外侵襲實驗,檢測PCDGF反義RNA真覈錶達載體對Sw626和A2780細胞增殖和侵襲能力的影響.採用Western blot技術,檢測轉染PCDGF反義RNA真覈錶達載體前後Sw626細胞cyclin D1和CDK4蛋自錶達的變化.採用逆轉錄聚閤酶鏈反應(RT-PCR)和明膠酶譜法,分析PCDGF反義RNA真覈錶達載體對Sw626細胞基質金屬蛋白酶2(MMP-2)錶達和活性的影響.結果 與空白對照組相比,PCDGF反義覈痠載體轉染組Sw626和A2780細胞的增殖抑製率分彆為72.9%和70.9%,侵襲能力分彆被抑製瞭62.9%和59.0%.轉染組Sw626細胞cyclin D1和CDK4蛋白的錶達水平分彆為0.38±0.08和0.37±0.13,明顯低于空白對照組(0.84±0.11和0.64±0.11,P<0.01).與空白對照組(0.89±0.09)相比,轉染組Sw626細胞MMP-2 mRNA的錶達水平(0.66±0.11)雖未見降低(P>0.05),但MMP-2酶原的活性被叨顯抑製.結論 PCDGF反義覈痠可顯著抑製高度噁性人卵巢癌細胞株Sw626和A2780的增殖和侵襲能力,併逆轉其部分噁性錶型,這可能與其能下調cyclin D1和CDK4蛋白的錶達併抑製MMP-2酶原的活性有關;PCDGF可以作為卵巢癌治療的新靶點.
목적 관찰기태류세포원성생장인자(PCDGF)반의핵산재체대고도악성인란소암세포주Sw626화A2780증식화침습적억제효응,병초보탐토기상관궤제.방법 채용이분기추화사담서람(MTT)법화Boyden소질체외침습실험,검측PCDGF반의RNA진핵표체재체대Sw626화A2780세포증식화침습능력적영향.채용Western blot기술,검측전염PCDGF반의RNA진핵표체재체전후Sw626세포cyclin D1화CDK4단자표체적변화.채용역전록취합매련반응(RT-PCR)화명효매보법,분석PCDGF반의RNA진핵표체재체대Sw626세포기질금속단백매2(MMP-2)표체화활성적영향.결과 여공백대조조상비,PCDGF반의핵산재체전염조Sw626화A2780세포적증식억제솔분별위72.9%화70.9%,침습능력분별피억제료62.9%화59.0%.전염조Sw626세포cyclin D1화CDK4단백적표체수평분별위0.38±0.08화0.37±0.13,명현저우공백대조조(0.84±0.11화0.64±0.11,P<0.01).여공백대조조(0.89±0.09)상비,전염조Sw626세포MMP-2 mRNA적표체수평(0.66±0.11)수미견강저(P>0.05),단MMP-2매원적활성피도현억제.결론 PCDGF반의핵산가현저억제고도악성인란소암세포주Sw626화A2780적증식화침습능력,병역전기부분악성표형,저가능여기능하조cyclin D1화CDK4단백적표체병억제MMP-2매원적활성유관;PCDGF가이작위란소암치료적신파점.
Objective To investigate the inhibitory effects of an antisense PC cell derived growth factor (PCDGF) vector on proliferation and invasion of highly malignant ovarian cancer cell lines Sw626 and A2780 cells, and preliminarily explore the related mechanisms. Methods MTT assay and Boyden chamber in vitro invasion assay were employed to detect the changes of proliferation and invasion ability in the Sw626 and A2780 cells transfected with anti-sense PCDGF. The expression levels of cyclin D1 and CDK4 proteins before and after transfection were detected by Western blotting. The effects on the expression and activity of MMP-2 were evaluated by quantitative RT-PCR and zymography,respectively. Results Comparing with the blank group, the proliferation inhibition rate of the Sw626 and A2780 cells transfected with anti-sense PCDGF was 72.9% and 70.9%, respectively, and the invasion ability was inhibited by 62.9% and 59.0%, respectively. The levels of cyclin D1 and CDK4 protein expression in antisense PCDGF transfected cells were 0.38±0.08 and 0.37±0.13, respectively, all significantly lower than 0.84±0.11 and 0.64±0.11, respectively, in the blank group (P<0.01). The MMP-2 mRNA expression level in antisense PCDGF transfected cell group was 0.66±0.11, not significantly decreased in comparison with 0.89±0.09 in the blank group (P>0.05), but the activity of MMP-2 was inhibited significantly. Conclusion The antisense PCDGF vector may inhibit markedly the proliferation and invasion of highly malignant ovarian cancer cells, and partially reverses their malignant phenotype. It seems to be related with down-regulating the expression of cyclin DI and CDK4 and inhibiting the activity of MMP-2. Our findings indicate that PCDGF may become a new target for antisense gene therapy of ovarian cancer.
