中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
9期
3892-3896
,共5页
RNA,小分子干扰%RIP1基因%结直肠肿瘤
RNA,小分子榦擾%RIP1基因%結直腸腫瘤
RNA,소분자간우%RIP1기인%결직장종류
RNA,small Interfering%RIP1 gene%Colorectal neoplasms
目的探讨 RNA 干扰沉默 RIP1基因表达对人大肠癌 Lovo 细胞生物学行为的影响。方法将人大肠癌Lovo细胞常规分为空白对照、阴性对照组和实验转染组。体外化学合成RIP1基因序列特异性小干扰RNA（siRNA），Hegene介导转染人大肠癌细胞株Lovo细胞；采用逆转录-聚合酶链反应（RT-PCR）方法检测特异性siRNA对RIP1基因在mRNA水平上沉默效果；转染后采用噻唑蓝（ MrI′r）比色法检测siRNA对细胞增殖的影响；流式细胞周期法检测siRNA对细胞周期的影响；Transwell试验体外检测细胞迁移和侵袭能力。结果成功转染RIP1 siRNA的大肠癌Lovo细胞，RT-PCR显示RIP1 mRNA和蛋白表达在相应的癌细胞中明显下降；与阴性对照组比较，细胞增殖明显抑制（ P ＜0.05）；G0～G1期细胞[（54.68±1.54）％]明显多于阴性对照组[（48.48±1.96）％]和空白对照组[（43.92±1.68）％]；RNA干扰明显抑制Lovo细胞迁移力和侵袭力（21±2.731 vs．43±2.064）。结论 RIP1 siRNA可有效抑制大肠癌Lovo细胞RIP1基因的表达，从而抑制肿瘤细胞增殖，促进细胞凋亡，抑制细胞迁移和侵袭，RIP1 siRNA能有效调控Lovo细胞恶性生物学行为。
目的探討 RNA 榦擾沉默 RIP1基因錶達對人大腸癌 Lovo 細胞生物學行為的影響。方法將人大腸癌Lovo細胞常規分為空白對照、陰性對照組和實驗轉染組。體外化學閤成RIP1基因序列特異性小榦擾RNA（siRNA），Hegene介導轉染人大腸癌細胞株Lovo細胞；採用逆轉錄-聚閤酶鏈反應（RT-PCR）方法檢測特異性siRNA對RIP1基因在mRNA水平上沉默效果；轉染後採用噻唑藍（ MrI′r）比色法檢測siRNA對細胞增殖的影響；流式細胞週期法檢測siRNA對細胞週期的影響；Transwell試驗體外檢測細胞遷移和侵襲能力。結果成功轉染RIP1 siRNA的大腸癌Lovo細胞，RT-PCR顯示RIP1 mRNA和蛋白錶達在相應的癌細胞中明顯下降；與陰性對照組比較，細胞增殖明顯抑製（ P ＜0.05）；G0～G1期細胞[（54.68±1.54）％]明顯多于陰性對照組[（48.48±1.96）％]和空白對照組[（43.92±1.68）％]；RNA榦擾明顯抑製Lovo細胞遷移力和侵襲力（21±2.731 vs．43±2.064）。結論 RIP1 siRNA可有效抑製大腸癌Lovo細胞RIP1基因的錶達，從而抑製腫瘤細胞增殖，促進細胞凋亡，抑製細胞遷移和侵襲，RIP1 siRNA能有效調控Lovo細胞噁性生物學行為。
목적탐토 RNA 간우침묵 RIP1기인표체대인대장암 Lovo 세포생물학행위적영향。방법장인대장암Lovo세포상규분위공백대조、음성대조조화실험전염조。체외화학합성RIP1기인서렬특이성소간우RNA（siRNA），Hegene개도전염인대장암세포주Lovo세포；채용역전록-취합매련반응（RT-PCR）방법검측특이성siRNA대RIP1기인재mRNA수평상침묵효과；전염후채용새서람（ MrI′r）비색법검측siRNA대세포증식적영향；류식세포주기법검측siRNA대세포주기적영향；Transwell시험체외검측세포천이화침습능력。결과성공전염RIP1 siRNA적대장암Lovo세포，RT-PCR현시RIP1 mRNA화단백표체재상응적암세포중명현하강；여음성대조조비교，세포증식명현억제（ P ＜0.05）；G0～G1기세포[（54.68±1.54）％]명현다우음성대조조[（48.48±1.96）％]화공백대조조[（43.92±1.68）％]；RNA간우명현억제Lovo세포천이력화침습력（21±2.731 vs．43±2.064）。결론 RIP1 siRNA가유효억제대장암Lovo세포RIP1기인적표체，종이억제종류세포증식，촉진세포조망，억제세포천이화침습，RIP1 siRNA능유효조공Lovo세포악성생물학행위。
Objective To study the effect of small interference RNA ( siRNA ) silencing RIP1 on the biological behavior of human colon cancer cell line Lovo .Methods Lovo cells were divided into three groups:the bland control group,the negative control group,the RIP1-siRNA group.Chemically synthesized siRNA targeting RIP1 (RIP1-siRNA) was transfected into Lovo cells with high metastatic potential by Hygene .Reverse transcription-polymerase chain reaction(RT-PCR)was used to semi-quantify the RIP1 mRNA level.Then proliferation of Lovo cells was determined by methyl thiazol tetrazolium ( MTT) assay.Flow cytometry was used for cell cycle analysis .The activities of motility and invasion of Lovo cells were assessed by transwell chamber inwasion assay in vitro, respectively.Results In the RIP1-siRNA group,the RIP1 mRNA level was down-regulaed remarkably(P<0.05), the abilities of proliferation were inhibited ,the number of cells in G0-G1 phrase increased in RIP1-siRNA group [(54.68 ±1.54)%] in comparison with negative control [(48.48 ±1.96)%] and blank control groups [(43.92 ±1.68)%],the motility and inwation of Lovo cells were inhibited significantly (21 ±2.731 vs.43 ±2.064). Conclusions Silencing RIP1 could regulate the malignant biological behavrors of colon cancer cell line Lovo effectively.
