HOXA13在非小细胞肺癌组织中的表达及其对A549细胞和移植瘤生长的影响
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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。q PCR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,q PCR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 m RNA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。
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.
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.
引文
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[11]单艳,李志刚,姬卫国,等.VEGFR2基因V297I位点对贝伐珠单抗联合化疗一线治疗晚期非小细胞肺癌患者临床疗效的影响[J].中国肿瘤生物治疗杂志,2019,26(1):67-72.DOI:10.3872/j.issn.1007-385x.2019.01.011.
[12]曲宝亮,穆怀博,勾建强,等.lncRNA HCG18/miR-17-5p/HMGA2分子轴调控非小细胞肺癌细胞增殖及迁移[J].中国肿瘤生物治疗杂志,2019,26(4):409-416.10.3872/j.issn.1007-385X.2019.04.007.
[13]戴素丽,白函瑜,王耀杰,等.非小细胞肺癌组织中高表达的miR-1269对肺癌细胞A549生物学行为的影响[J].中国肿瘤生物治疗杂志,2018,25(12):1282-1289.DOI:10.3872/j.issn.1007-385x.2018.12.012.
[14]MA R L,SHEN L Y,CHEN K N.Coexpression of ANXA2,SOD2and HOXA13 predicts poor prognosis of esophageal squamous cell carcinoma[J].Oncol Rep,2014,31(5):2157-2164.DOI:10.3892/or.2014.3088.
[15]QUAGLIATA L,QUINTAVALLE C,LANZAFAME M,et al.High expression of HOXA13 correlates with poorly differentiated hepatocellular carcinomas and modulates sorafenib response in in vitro models[J].Lab Invest,2018,98(1):95-105.DOI:10.1038/labinvest.2017.107.
[16]QU L P,ZHONG Y M,ZHENG Z,et al.CDH17 is a downstream effector of HOXA13 in modulating the Wnt/β-catenin signaling pathway in gastric cancer[J].Eur Rev Med Pharmacol Sci,2017,21(6):1234-1241.
[17]李欢,王梦杰,张翔宇,等.桥接整合因子1通过c-MYC途径抑制非小细胞肺癌A549细胞中PD-L1的表达[J].中国肿瘤生物治疗杂志,2018,25(8):762-766.DOI:10.3872/j.issn.1007-385x.2018.08.002.
[18]蔡华荣,王志强,江跃全.长链非编码RNA UCA1靶向调控miR-185-5p对非小细胞肺癌A549细胞的作用及其机制[J].中国肿瘤生物治疗杂志,2018,25(6):555-561.DOI:10.3872/j.issn.1007-385x.2018.06.002.
[2]杜凤华,闵旭红,梅晓冬.肺腺癌靶向治疗药物的应用及耐药机制[J].临床肺科杂志,2018,23(1):168-172.DOI:10.3969/j.issn.1009-6663.2018.01.046.
[3]JUNG C Y,ANTONIA S J.Tumor immunology and immune checkpoint inhibitors in non-small cell lung cancer[J/OL].Tuberc Respir Dis(Seoul),2018,81(1):29-41[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5771744/.DOI:10.4046/trd.2017.0120.
[4]OKAFOR M T,NWAGHA T U,ANUSIEM C,et al.Cancer prevention,the need to preserve the integrity of the genome at all cost[J].Niger J Clin Pract,2018,21(5):539-545.DOI:10.4103/njcp.njcp_272_17.
[5]CAO L H,CHEN C,LENG Y J,et al.A missense mutation of HOXA13 underlies hand-foot-genital syndrome in a Chinese family[J].J Genet,2017,96(4):647-652.
[6]WEN Y A,SHU F P,CHEN Y D,et al.The prognostic value of HOXA13 in solid tumors:A meta-analysis[J].Clin Chim Acta,2018,483:64-68.DOI:10.1016/j.cca.2018.04.024.
[7]李霞,汤志远,冯健.Rab27A蛋白表达在非小细胞肺癌中的研究进展[J].南通大学学报(医学版),2017,37(1):58-62.DOI:10.16424/j.cnki.cn32-1807/r.2017.01.015.
[8]张艳苗,顾玉海.低氧微环境与非小细胞肺癌关系的研究进展[J].临床肺科杂志,2017,22(4):746-749.DOI:10.3969/j.issn.1009-6663.2017.04.047.
[9]林志川,王飞,陈谭根.同源异型盒基因调控肝细胞肝癌的生物学机制[J].中华实验外科杂志,2016,33(5):1218-1221.DOI:10.3760/cma.j.issn.1001-9030.2016.05.015.
[10]HAN Y,SONG C L,WANG J Y,et al.HOXA13 contributes to gastric carcinogenesis through DHRS2 interacting with MDM2 and confers 5-FU resistance by a p53-dependent pathway[J].Mol Carcinog,2018,57(6):722-734.DOI:10.1002/mc.22793.
[11]单艳,李志刚,姬卫国,等.VEGFR2基因V297I位点对贝伐珠单抗联合化疗一线治疗晚期非小细胞肺癌患者临床疗效的影响[J].中国肿瘤生物治疗杂志,2019,26(1):67-72.DOI:10.3872/j.issn.1007-385x.2019.01.011.
[12]曲宝亮,穆怀博,勾建强,等.lncRNA HCG18/miR-17-5p/HMGA2分子轴调控非小细胞肺癌细胞增殖及迁移[J].中国肿瘤生物治疗杂志,2019,26(4):409-416.10.3872/j.issn.1007-385X.2019.04.007.
[13]戴素丽,白函瑜,王耀杰,等.非小细胞肺癌组织中高表达的miR-1269对肺癌细胞A549生物学行为的影响[J].中国肿瘤生物治疗杂志,2018,25(12):1282-1289.DOI:10.3872/j.issn.1007-385x.2018.12.012.
[14]MA R L,SHEN L Y,CHEN K N.Coexpression of ANXA2,SOD2and HOXA13 predicts poor prognosis of esophageal squamous cell carcinoma[J].Oncol Rep,2014,31(5):2157-2164.DOI:10.3892/or.2014.3088.
[15]QUAGLIATA L,QUINTAVALLE C,LANZAFAME M,et al.High expression of HOXA13 correlates with poorly differentiated hepatocellular carcinomas and modulates sorafenib response in in vitro models[J].Lab Invest,2018,98(1):95-105.DOI:10.1038/labinvest.2017.107.
[16]QU L P,ZHONG Y M,ZHENG Z,et al.CDH17 is a downstream effector of HOXA13 in modulating the Wnt/β-catenin signaling pathway in gastric cancer[J].Eur Rev Med Pharmacol Sci,2017,21(6):1234-1241.
[17]李欢,王梦杰,张翔宇,等.桥接整合因子1通过c-MYC途径抑制非小细胞肺癌A549细胞中PD-L1的表达[J].中国肿瘤生物治疗杂志,2018,25(8):762-766.DOI:10.3872/j.issn.1007-385x.2018.08.002.
[18]蔡华荣,王志强,江跃全.长链非编码RNA UCA1靶向调控miR-185-5p对非小细胞肺癌A549细胞的作用及其机制[J].中国肿瘤生物治疗杂志,2018,25(6):555-561.DOI:10.3872/j.issn.1007-385x.2018.06.002.