下调叉头转录因子M1基因表达对非小细胞肺癌细胞生长与侵袭能力的影响

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.

Role of inhibition of transcription factor fockhead box M1 expression by RNA interference in the proliferation and invasiveness of non-small cell lung cancer cells

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.