浙江医学
浙江醫學
절강의학
ZHEJIANG MEDICAL JOURNAL
2014年
5期
378-381
,共4页
蔡炜龙%汪伟民%潘杰%王雁%许洪宝%马志红
蔡煒龍%汪偉民%潘傑%王雁%許洪寶%馬誌紅
채위룡%왕위민%반걸%왕안%허홍보%마지홍
胃癌%ATDC%RNA干扰%增殖
胃癌%ATDC%RNA榦擾%增殖
위암%ATDC%RNA간우%증식
Gastric cancer%ATDC%siRNA%Proliferation
目的:通过比较ataxia- telangiectasia group D complementing gene(ATDC)在永生化胃黏膜细胞GES与胃癌细胞系MNK45、AGS中的表达差异,探讨其对胃癌细胞生长、增殖及侵袭能力的影响。方法应用Western blot实验检测ATDC在胃癌细胞系MKN45、AGS及永生化胃黏膜细胞系GES表达差异,利用慢病毒携带的siRNA下调ATDC在MKN45细胞表达水平,采用MTT实验检测细胞生长能力、流式细胞术检测细胞周期、平板克隆实验检测细胞克隆形成能力以及transwel 实验检测细胞侵袭能力。结果 ATDC在人胃癌细胞系MKN45与AGS细胞中的表达水平均明显高于永生化胃黏膜细胞GES(均P<0.05);转染siRNA慢病毒后,与Con- MKN45及MKN45比较,Si- MKN45细胞的表达水平明显下降(P<0.05);下调ATDC表达后MKN45细胞的生长能力明显受到抑制(P<0.05),S期细胞明显减少、增殖指数克隆形成率及侵袭能力均明显降低(均P<0.05)。结论慢病毒携带的siRNA下调ATDC表达后能够抑制胃癌细胞MKN45的生长、增殖与侵袭能力,可为胃癌的基因治疗提供新的靶点。
目的:通過比較ataxia- telangiectasia group D complementing gene(ATDC)在永生化胃黏膜細胞GES與胃癌細胞繫MNK45、AGS中的錶達差異,探討其對胃癌細胞生長、增殖及侵襲能力的影響。方法應用Western blot實驗檢測ATDC在胃癌細胞繫MKN45、AGS及永生化胃黏膜細胞繫GES錶達差異,利用慢病毒攜帶的siRNA下調ATDC在MKN45細胞錶達水平,採用MTT實驗檢測細胞生長能力、流式細胞術檢測細胞週期、平闆剋隆實驗檢測細胞剋隆形成能力以及transwel 實驗檢測細胞侵襲能力。結果 ATDC在人胃癌細胞繫MKN45與AGS細胞中的錶達水平均明顯高于永生化胃黏膜細胞GES(均P<0.05);轉染siRNA慢病毒後,與Con- MKN45及MKN45比較,Si- MKN45細胞的錶達水平明顯下降(P<0.05);下調ATDC錶達後MKN45細胞的生長能力明顯受到抑製(P<0.05),S期細胞明顯減少、增殖指數剋隆形成率及侵襲能力均明顯降低(均P<0.05)。結論慢病毒攜帶的siRNA下調ATDC錶達後能夠抑製胃癌細胞MKN45的生長、增殖與侵襲能力,可為胃癌的基因治療提供新的靶點。
목적:통과비교ataxia- telangiectasia group D complementing gene(ATDC)재영생화위점막세포GES여위암세포계MNK45、AGS중적표체차이,탐토기대위암세포생장、증식급침습능력적영향。방법응용Western blot실험검측ATDC재위암세포계MKN45、AGS급영생화위점막세포계GES표체차이,이용만병독휴대적siRNA하조ATDC재MKN45세포표체수평,채용MTT실험검측세포생장능력、류식세포술검측세포주기、평판극륭실험검측세포극륭형성능력이급transwel 실험검측세포침습능력。결과 ATDC재인위암세포계MKN45여AGS세포중적표체수평균명현고우영생화위점막세포GES(균P<0.05);전염siRNA만병독후,여Con- MKN45급MKN45비교,Si- MKN45세포적표체수평명현하강(P<0.05);하조ATDC표체후MKN45세포적생장능력명현수도억제(P<0.05),S기세포명현감소、증식지수극륭형성솔급침습능력균명현강저(균P<0.05)。결론만병독휴대적siRNA하조ATDC표체후능구억제위암세포MKN45적생장、증식여침습능력,가위위암적기인치료제공신적파점。
Objective To investigate the effect of ATDC gene silencing by siRNA on the proliferation and migration of gastric cancer cells. Methods The expression of ataxia- telangiectasia group D complementing(ATDC) was detected in immor-talized gastric mucosal epithelium cellline GES and gastric cancer celllines MKN45, AGS by Western blot. Lentivirus- mediated siRNA targeting ATDC was transfected into MKN45 cells, and western blot was performed to evaluate the inhibitory effect of siR-NA on ATDC expression. The proliferation, cellcycle, growth potentiality and migration of MKN45 cells were evaluated by MTT, plate cloning formation test, flow cytometry and transwel assays, respectively. Results The expression of ATDC was signifi-cantly higher in both MKN45 and AGS cells compared with GES cells. Lentivirus- mediated siRNA targeting ATDC markedly re-pressed the expression of ATDC in MKN45 cells. MTT and plate cloning formation assays showed that down- regulation of ATDC significantly inhibited the proliferation and growth of MKN45 cells (P<0.05). Flow cytometry showed that the number of cells transfected with ATDC siRNA had higher G1, lower S phase and lower proliferation index (PI) (P<0.05) than those of controls. Transwel assays revealed that the migration of cells transfected with ATDC siRNA was significantly deceased compared with controls. Conclusion The expression of ATDC is increased in gastric cancer cells, and silencing of ATDC significantly sup-presses the proliferation and migration of gastric cancer cells, indicating that ATDC may be a novel target for gene therapy of gastric cancer.