中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2009年
34期
2424-2428
,共5页
周磊%张萍海%徐欣%许诺%高磊%白春学%张新
週磊%張萍海%徐訢%許諾%高磊%白春學%張新
주뢰%장평해%서흔%허낙%고뢰%백춘학%장신
癌%非小细胞肺%叉头转录因子类%细胞增殖%肿瘤侵润%RNA干扰
癌%非小細胞肺%扠頭轉錄因子類%細胞增殖%腫瘤侵潤%RNA榦擾
암%비소세포폐%차두전록인자류%세포증식%종류침윤%RNA간우
Carcinoma%non-small-cell lung%Fockhead transcription factors%Cell proliferation%Neoplasm invasiveness%RNA interference
目的 研究用叉头转录因子M1(FoxM1)基因小干扰RNA下调非小细胞肺癌(NSCLC)细胞株FoxM1基因表达后细胞增殖与侵袭能力的改变,为开发新的NSCLC靶向治疗方法提供依据.方法 设计靶向FoxM1小干扰RNA(siFoxM1),分别以siFoxM1和无关对照siRNA转染人NSCLC细胞株SPC-A-1、A549和LTEP-a-2,采用实时逆转录PCR和蛋白质印迹法检测FoxM1 mRNA和蛋白表达,采用集落形成试验、划痕试验和细胞侵袭试验研究下调FoxM1基因表达对细胞增殖、迁移和侵袭能力的影响.结果 转染siFoxM1可高效特异地下调SPC-A-1、A549和LTEP-a-2细胞的FoxM1mRNA表达(分别下降83.9%、83.6%、88.6%)和蛋白表达.转染siFoxM1的SPC-A-1、A549和LTEP-a-2细胞集落形成数[分别为(136.0±15.5)、(87.0±2.6)和(121.7±9.4)个]和穿膜细胞数[分别为(19.2±2.5)、(4.2±0.8)和(6.2±1.8)个]均明显低于转染无关对照siRNA细胞[集落形成数分别为(222.3±20.5)、(164.7±14.1)和(260.7±13.5)个,穿膜细胞数分别为(81.4±6.2)、(39.2±4.6)和(35.6±3.0)个,均P<0.01];转染siFoxM1的SPC-A-1细胞划痕愈合率(52.6%±7.8%)明显低于转染无关对照siRNA细胞(85.3%±18.6%,P<0.01).结论 FoxM1基因表达下调可显著降低多种NSCLC细胞的增殖与侵袭能力,提爪FoxM1是潜在的肺癌治疗靶点.
目的 研究用扠頭轉錄因子M1(FoxM1)基因小榦擾RNA下調非小細胞肺癌(NSCLC)細胞株FoxM1基因錶達後細胞增殖與侵襲能力的改變,為開髮新的NSCLC靶嚮治療方法提供依據.方法 設計靶嚮FoxM1小榦擾RNA(siFoxM1),分彆以siFoxM1和無關對照siRNA轉染人NSCLC細胞株SPC-A-1、A549和LTEP-a-2,採用實時逆轉錄PCR和蛋白質印跡法檢測FoxM1 mRNA和蛋白錶達,採用集落形成試驗、劃痕試驗和細胞侵襲試驗研究下調FoxM1基因錶達對細胞增殖、遷移和侵襲能力的影響.結果 轉染siFoxM1可高效特異地下調SPC-A-1、A549和LTEP-a-2細胞的FoxM1mRNA錶達(分彆下降83.9%、83.6%、88.6%)和蛋白錶達.轉染siFoxM1的SPC-A-1、A549和LTEP-a-2細胞集落形成數[分彆為(136.0±15.5)、(87.0±2.6)和(121.7±9.4)箇]和穿膜細胞數[分彆為(19.2±2.5)、(4.2±0.8)和(6.2±1.8)箇]均明顯低于轉染無關對照siRNA細胞[集落形成數分彆為(222.3±20.5)、(164.7±14.1)和(260.7±13.5)箇,穿膜細胞數分彆為(81.4±6.2)、(39.2±4.6)和(35.6±3.0)箇,均P<0.01];轉染siFoxM1的SPC-A-1細胞劃痕愈閤率(52.6%±7.8%)明顯低于轉染無關對照siRNA細胞(85.3%±18.6%,P<0.01).結論 FoxM1基因錶達下調可顯著降低多種NSCLC細胞的增殖與侵襲能力,提爪FoxM1是潛在的肺癌治療靶點.
목적 연구용차두전록인자M1(FoxM1)기인소간우RNA하조비소세포폐암(NSCLC)세포주FoxM1기인표체후세포증식여침습능력적개변,위개발신적NSCLC파향치료방법제공의거.방법 설계파향FoxM1소간우RNA(siFoxM1),분별이siFoxM1화무관대조siRNA전염인NSCLC세포주SPC-A-1、A549화LTEP-a-2,채용실시역전록PCR화단백질인적법검측FoxM1 mRNA화단백표체,채용집락형성시험、화흔시험화세포침습시험연구하조FoxM1기인표체대세포증식、천이화침습능력적영향.결과 전염siFoxM1가고효특이지하조SPC-A-1、A549화LTEP-a-2세포적FoxM1mRNA표체(분별하강83.9%、83.6%、88.6%)화단백표체.전염siFoxM1적SPC-A-1、A549화LTEP-a-2세포집락형성수[분별위(136.0±15.5)、(87.0±2.6)화(121.7±9.4)개]화천막세포수[분별위(19.2±2.5)、(4.2±0.8)화(6.2±1.8)개]균명현저우전염무관대조siRNA세포[집락형성수분별위(222.3±20.5)、(164.7±14.1)화(260.7±13.5)개,천막세포수분별위(81.4±6.2)、(39.2±4.6)화(35.6±3.0)개,균P<0.01];전염siFoxM1적SPC-A-1세포화흔유합솔(52.6%±7.8%)명현저우전염무관대조siRNA세포(85.3%±18.6%,P<0.01).결론 FoxM1기인표체하조가현저강저다충NSCLC세포적증식여침습능력,제조FoxM1시잠재적폐암치료파점.
Objective To investigate the change of proliferation and invasiveness of non-small cell lung cancer (NSCLC) cell lines SPC-A-1, A549 and LTEP-a-2 with fockhead box M1 (FoxM1) expression deficiency. Methods A siRNA targeting FoxM1 was designed to deplete the FoxM1 expression of these cell lines and an unrelated siRNA used as control. Real-time RT-PCR and Western blotting were used to examine the FoxM1 expression in mRNA and protein level respectively. Colony assay, wound healing assay and transwell chamber assay were employed to evaluate the colony formation ability and invasiveness of FoxM1 deficient cells. Results The designed siRNA could efficiently deplete FoxM1 expression by 83.9%, 83.6% and 88.6% in SPC-A-1, A549 and LTEP-a-2 cell lines respectively. Real-time RT-PCR and Western blot test showed that the FoxM1 protein was also depleted. The colony formation numbers (136.0± 15.5, 87.0±2.6 and 121.7±9.4 respectively) and invasion cell numbers (19.2±2.5, 4.2±0.8 and 6.2±1.8 respectively) of FoxM1 deficient SPC-A-1, A549 and LTEP-a-2 cell lines were significantly fewer than those of the unrelated-siRNA transfected group (colony formation numbers were 222.3±20.5, 164.7±14.1 and 260.7±13.5 respectively, and invasive cell numbers were 81.4±6.2, 39.2±4.6 and 35.6±3.0 respectively, all P<0.01). The cell migration rate of siFoxM1 deficient SPC-A-1 (52.6%± 7.8%) was significantly lower than that of the unrelated-siRNA transfected group (85.3%±18.6% ,P < 0.01). Conclusions The proliferation and invasiveness of several NSCLC cell lines were significantly inhibited after the FoxM1 gene expression was depleted. It suggests that inhibiting the FoxM1 expression might be a promising way for lung cancer therapy.
