同源盒A13在胃癌组织中表达对SGC-7901细胞增殖和侵袭的影响

Influence of homeobox gene A13 in gastric cancer on proliferation and invasion of SGC-7901 cells

目的探讨胃癌组织及癌旁组织中同源盒A13(homeobox gene A13,HOXA13)表达差异,及下调胃癌SGC-7901细胞HOXA13基因表达对细胞增殖和侵袭能力的影响。方法取109例胃癌患者手术切除的胃癌组织和癌旁正常组织,采用免疫组织化学法检测HOXA13蛋白表达;取对数生长期SGC-7901细胞分为转染组(转染siRNA-HOXA13)、阴性对照组(转染siRNA-NC)和空白对照组(不作任何处理),采用MTT法检测转染12、24、48、72、96h时细胞增殖能力,Transwell法检测转染后培养48h时细胞侵袭能力,Western blot法检测转染后培养48h时细胞HOXA13、Twist、E-cadherin和Vimentin蛋白表达。结果胃癌组织HOXA13蛋白阳性表达率(75.23%)高于癌旁正常组织(33.94%)(P<0.05);HOXA13蛋白阳性表达率在进展期胃癌(82.98%)、低分化程度(84.93%)、TNM分期Ⅲ～Ⅳ期(86.27%)、有淋巴结转移者(84.38%)均高于早期胃癌(26.67%)、中高分化程度(55.56%)、TNM分期Ⅰ～Ⅱ期(65.52%)、无淋巴结转移者(62.22%)(P<0.05),在性别、年龄、肿瘤直径上差异均无统计学意义(P>0.05);转染组转染后培养24、48、72、96h时吸光度值低于阴性对照组和空白对照组(P<0.05),阴性对照组与空白对照组比较差异无统计学意义(P>0.05);转染组转染后培养48h时侵袭细胞数[(97.14±10.03)个]均较阴性对照组[(115.39±5.77)个]和空白对照组[(125.11±12.45)个]少(P<0.05),阴性对照组与空白对照组比较差异无统计学意义(P>0.05);转染组转染后培养48h时细胞HOXA13、Twist和Vimentin蛋白表达(0.24±0.05、0.34±0.07、0.36±0.04)均低于阴性对照组(0.87±0.11、0.75±0.02、0.75±0.07)和空白对照组(0.88±0.05、0.76±0.05、0.72±0.06)(P<0.05),E-cadherin蛋白表达(0.67±0.08)高于阴性对照组(0.33±0.06)和空白对照组(0.35±0.04)(P<0.05),阴性对照组与空白对照组比较差异无统计学意义(P>0.05)。结论胃癌组织中HOXA13蛋白呈高表达,其表达程度与肿瘤恶性程度有关;下调SGC-7901细胞HOXA13基因表达可抑制细胞增殖和侵袭能力。

Objective To investigate the expression of homeobox gene A13（HOXA13）in gastric cancer tissues and its adjacent normal tissues,and to explore the influence of down-regulation of HOXA13 gene expression on cell proliferation and invasion of gastric cancer SGC-7901 cells. Methods The expression of HOXA13 protein was detected by immunohistochemical method in gastric cancer tissues and its adjacent normal tissues from 109 patients with gastric cancer.SGC-7901 cells in logarithmic phase were divided into transfection group transfected with siRNA-HOXA13,negative control group transfected with siRNA-NC and blank control group with no treatment.MTT assay was used to detect cell proliferations in 12,24,48,72 and 96 hafter transfection.Transwell assay was used to detect cell invasion in48 hafter transfection.Western blot was used to detect the expressions of HOXA13,Twist,E-cadherin and Vimentin proteins in cells in 48 hafter transfection.Results The positive expression rate of HOXA13 protein was significantly higher in gastric cancer tissues（75.23%）than that in adjacent normal tissues（33.94%）（P〈0.05）.The positive expression rate of HOXA13 protein was significantly higher in patients with advanced gastric cancer,poorly differentiation,TNM stage Ⅲ-Ⅳ and lymph node metastasis（82.98%,84.93%,86.27%,84.38%）than that in patients with early gastric cancer,moderately differentiation,TNM stageⅠ-Ⅱand no lymph node metastasis（26.67%,55.56%,65.52%,62.22%）（P〈0.05）,and there were no significant differences in the gender,age and tumor diameter（P〉0.05）.The optical density was significantly lower in transfection group in 24,48,72 and 96 hafter transfection than that in negative control group and blank control group（P〈0.05）,and there was no significant difference between negative control group and blank control group（P〉0.05）.The number of invasive cells was significantly lower in transfection group（97.14±10.03）than that in negative control group（115.39±5.77）and blank control group（125.11±12.45）（P〈0.05）,and there was no significant difference between negative control group and blank control group（P〉0.05）.The relative expressions of HOXA13,Twist and Vimentin proteins were significantly lower in transfection group（0.24±0.05,0.34±0.07,0.36±0.04）than those in negative control group（0.87±0.11,0.75±0.02,0.75±0.07）and blank control group（0.88±0.05,0.76±0.05,0.72±0.06）in 48 hafter transfection（P〈0.05）,the relative expression of E-cadherin protein was significantly higher in transfection group（0.67±0.08）than that in negative control group（0.33±0.06）and blank control group（0.35±0.04）（P〈0.05）,and there was no significant difference between negative control group and blank control group（P〉0.05）.Conclusion HOXA13 protein is highly expressed in gastric cancer tissues and is associated with tumor malignancies.Down-regulation of HOXA13 gene expression could effectively inhibit cell proliferation and invasion,probably by reversing epithelial-mesenchymal transition process.