中华胃肠外科杂志
中華胃腸外科雜誌
중화위장외과잡지
CHINESE JOURNAL OF GASTROINTESTINAL SURGERY
2011年
1期
61-64
,共4页
韩丁培%崔江涛%陆爱国%陈雪华%冯波%宗雅萍%瞿顺%曹奇峰%郑民华
韓丁培%崔江濤%陸愛國%陳雪華%馮波%宗雅萍%瞿順%曹奇峰%鄭民華
한정배%최강도%륙애국%진설화%풍파%종아평%구순%조기봉%정민화
结直肠肿瘤%Polo样激酶1%RNA干扰%细胞迁移%肿瘤侵润
結直腸腫瘤%Polo樣激酶1%RNA榦擾%細胞遷移%腫瘤侵潤
결직장종류%Polo양격매1%RNA간우%세포천이%종류침윤
Colorectal neoplasms%Polo-like kinase 1%RNA interference%Cell migration%Neoplasm invasion
目的 研究PLK1在结直肠癌细胞迁移和侵袭过程中的作用.方法 常规培养9株结直肠癌细胞,采用PCR和Western-blot法筛选PLK1高表达细胞株.设计3种RNA干扰片段,转染高表达PLK1的结直肠癌细胞株,利用实时定量PCR和western-blot法验证并干扰效果.选择高干扰效果的干扰片段,通过Transwe1l实验来评估转染后结直肠癌细胞迁移和侵袭能力的变化.结果 SW1116细胞中PLK1 mRNA及蛋白都呈现高表达;3种干扰片段转染SW1116细胞后,PLK1表达量均有不同程度下降,以片段1干扰效果最为明显.在迁移实验中,PLK1干扰组穿膜细胞数为(44±14)个,低于阴性对照组[(242±40)个]和空白对照组[(240±38)个];在侵袭实验中,PLK1干扰组穿膜细胞数为(62±3)个,低于阴性对照组[(207±12)个]和空白对照组[(211±15)个];上述差异均有统计学意义(P＜0.01).结论 干扰PLK1能显著抑制肿瘤细胞的迁移、侵袭能力,提示PLK1可能在结直肠癌浸润和转移中具有一定作用.
目的 研究PLK1在結直腸癌細胞遷移和侵襲過程中的作用.方法 常規培養9株結直腸癌細胞,採用PCR和Western-blot法篩選PLK1高錶達細胞株.設計3種RNA榦擾片段,轉染高錶達PLK1的結直腸癌細胞株,利用實時定量PCR和western-blot法驗證併榦擾效果.選擇高榦擾效果的榦擾片段,通過Transwe1l實驗來評估轉染後結直腸癌細胞遷移和侵襲能力的變化.結果 SW1116細胞中PLK1 mRNA及蛋白都呈現高錶達;3種榦擾片段轉染SW1116細胞後,PLK1錶達量均有不同程度下降,以片段1榦擾效果最為明顯.在遷移實驗中,PLK1榦擾組穿膜細胞數為(44±14)箇,低于陰性對照組[(242±40)箇]和空白對照組[(240±38)箇];在侵襲實驗中,PLK1榦擾組穿膜細胞數為(62±3)箇,低于陰性對照組[(207±12)箇]和空白對照組[(211±15)箇];上述差異均有統計學意義(P＜0.01).結論 榦擾PLK1能顯著抑製腫瘤細胞的遷移、侵襲能力,提示PLK1可能在結直腸癌浸潤和轉移中具有一定作用.
목적 연구PLK1재결직장암세포천이화침습과정중적작용.방법 상규배양9주결직장암세포,채용PCR화Western-blot법사선PLK1고표체세포주.설계3충RNA간우편단,전염고표체PLK1적결직장암세포주,이용실시정량PCR화western-blot법험증병간우효과.선택고간우효과적간우편단,통과Transwe1l실험래평고전염후결직장암세포천이화침습능력적변화.결과 SW1116세포중PLK1 mRNA급단백도정현고표체;3충간우편단전염SW1116세포후,PLK1표체량균유불동정도하강,이편단1간우효과최위명현.재천이실험중,PLK1간우조천막세포수위(44±14)개,저우음성대조조[(242±40)개]화공백대조조[(240±38)개];재침습실험중,PLK1간우조천막세포수위(62±3)개,저우음성대조조[(207±12)개]화공백대조조[(211±15)개];상술차이균유통계학의의(P＜0.01).결론 간우PLK1능현저억제종류세포적천이、침습능력,제시PLK1가능재결직장암침윤화전이중구유일정작용.
Objective To examine the role of Polo-like kinase 1 (PLK1) in the migration and invasiveness of human colorectal cancer cells. Methods Nine colorectal cancer cell lines were cultured. Cell lines with the highest level of PLK1 expression was selected by PCR and Western blot. Three siRNA oligos segments targeting PLK1 were designed and selected cell lines transfected. Successful transfection was confirmed using real-time PCR and Western blot. Changes in migration and invasiveness of the selected cell line were evaluated by Transwell test. Results Colorectal cancer cell line SW1116 was selected with the highest expression of PLK1 at both mRNA level and protein level. The expression of PLK1 in SW1116 was reduced by the three siRNA oligos segments to varying degrees, and the No.l siRNA oligos segment was the most efficient. In migration test, the number of cells crossing through chambers in PLKI-siRNA group was 44±14, which was lower than that in the negative control group (242±40) and in blank control group(240±38). In invasion test,the number of cells crossing through chambers in PLK1-siRNA group was 62±3, which was lower than that in negative control group (207±12) and in blank control group (211±15). These differences were statistically significant (P<0.01). Conclusion PLK1 silencing by siRNA may inhibit the migration and invasiveness of colorectal cancer cells, suggesting that PLK1 might play an important role in the infiltration and metastasis of colorectal cancer.
