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[摘要]
[摘要] 目的:探讨HOXA13 在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的表达及沉默HOXA13 基因表达对A549 细胞和移植瘤生长的影响。方法:收集2014 年3 月至2016 年4 月在焦作煤业集团有限责任公司中央医院胸外科接受手术切除治疗的112 例NSCLC癌组织和相对应的癌旁组织。qPCR实验检测NSCLC癌和癌旁组织中HOXA13 表达。培养A549 细胞并分为siRNA-HOXA13 组、阴性对照组和对照组,qPCR实验检测A549 细胞中HOXA13 表达,CCK-8 法检测细胞增殖能力,Transwell法检测细胞侵袭能力。建立裸鼠移植瘤模型,观察裸鼠生长情况,5 周后处死,称瘤体质量并计算抑瘤率;qPCR实验检测瘤体组织中HOXA13 表达水平。结果:NSCLC 癌组织中HOXA13 mRNA相对表达量明显高于癌旁组织(1.83±0.13 vs 1.12±0.10,t=47.008,P=0.000),其相对表达量与TNM 分期、分化程度和淋巴结转移相关(P<0.05)。siRNA-HOXA13 组细胞中HOXA13 mRNA相对表达量低于阴性对照组和对照组(均P<0.05);siRNA-HOXA13 组24、48、72、96 h 时细胞增殖水平(D值)明显低于阴性对照组和对照组(F=30.727、5.427、13.816 和24.454,均P<0.05 或P<0.01);siRNA-HOXA13 组侵袭细胞数低于阴性对照组和对照组(均P<0.05);siRNA-HOXA13 组裸鼠移植瘤5 周时瘤体质量小于阴性对照组和对照组,而抑瘤率高于阴性对照组(均P<0.05);siRNA-HOXA13 组裸鼠移植瘤组织中HOXA13 mRNA相对表达量低于阴性对照组和对照组(均P<0.01)。结论:NSCLC癌组织中HOXA13 呈高表达,且与肿瘤发生、进展及转移有关;特异性沉默HOXA13 基因表达可抑制细胞增殖和侵袭力,并抑制裸鼠移植瘤生长。
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[Abstract]
[Abstract] Objective: To investigate the expression of HOXA13 in non-small cell lung cancer (NSCLC) tissues, and to explore the effect of silencing HOXA13 gene on the growth of A549 cells in vitro and xenograft in nude mice. Methods: A total of 112 pairs of NSCLC tissues and corresponding adjacent normal tissues from patients, who underwent surgical resection at the Department of Thoracic surgery, Central Hospital of Jiaozuo Coal Industry Group from March 2014 to April 2016, were selected for this study. Real-time fluorescence quantitative PCR was used to detect the expressions of HOXA13 in NSCLC tissues and adjacent tissues. A549 cells were cultured and divided into siRNA-HOXA13 group, negative control group and control group. Real-time quantitative PCR was used to detect the expression of HOXA13 in cells, CCK-8 was used to detect cell proliferation, and Transwell assay was used to detect cell invasion.Xenograft model in nude mice was constructed, and the growth of nude mice was observed. After 5 weeks, the mice was sacrificed and weighed, and the tumor-inhibition rate was calculated. Real-time fluorescence quantitative PCR(qPCR) was used to detect the expression of HOXA13 in xenograft tissues. Results: The relative expression level of HOXA13 mRNA in NSCLC tissues was significantly higher than that in the adjacent tissues (1.83±0.13 vs 1.12±0.10, t=47.008, P=0.000), and its expression was correlated to TNM staging,differentiation and lymph node metastasis (all P<0.05). The relative expression level of HOXA13 mRNA in cells of siRNAHOXA13 group was lower than that in the negative control group and the control group (P<0.05). The cell proliferation level (D values)at 24, 48, 72 and 96 h in the siRNA-HOXA13 group were significantly lower than those in the negative control group and control group (F=30.727, 5.427, 13.816 and 24.454, all P<0.05 or P<0.01); the number of invasive cells in the siRNA-HOXA13 group was lower than that in the negative control group and the control group (all P<0.05). The mass of xenograft in nude mice at week 5 in the siRNA-HOXA13 group was smaller than that in the negative control group and control group, while the tumor-inhibition rate was higher than the negative control group (all P<0.05). The relative mRNA expression level of HOXA13 in xenograft tumor tissue in the siRNA-HOXA13 group was lower than that in the negative control group and the control group (all P<0.01). Conclusion: HOXA13 was highly expressed in the non-small cell lung cancer tissues, and was related to oncogenesis, progression and metastasis of cancer. Specific silencing of HOXA13 gene expression could inhibit the proliferation and invasion of tumor cells and suppress the growth of xenograft in nude mice.
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[基金项目]
河南省科技发展计划资助项目(No.152300410158)