摘要
目的探讨TXNDC12在结直肠癌中的表达及其对结直肠癌细胞增殖、迁移和凋亡的影响。方法利用GEPIA和UALCAN在线网站预测TXNDC12在结直肠癌中的表达及其在淋巴结转移的结肠癌患者中的表达,并选取延安大学附属医院8例结直肠癌组织和癌旁组织通过Western blot检测TXNDC12蛋白表达水平。采用Sanger Box网站分析TXNDC12与结直肠癌组织免疫浸润的相关性。通过Western blot和RT-qPCR分别检测NCM460、HT-29、RKO和HCT116细胞中TXNDC12的蛋白和mRNA表达水平,利用小干扰RNA(small interfering RNA,siRNA)技术在HT-29和RKO细胞中敲低TXNDC12,并将其分为:空白对照组(MOCK组)、对照组(NC组)和敲低组(si-TXNDC12-2组),分别通过CCK-8实验和克隆形成实验、细胞划痕实验、Hochest荧光染色和Western blot检测结直肠癌细胞增殖能力、迁移能力以及细胞凋亡水平。结果GEPIA分析及临床病理组织标本Western blot结果显示,与正常组织相比,TXNDC12在结直肠癌组织中高表达(P<0.05);UALCAN分析结果显示TXNDC12在淋巴结转移的结肠癌患者中的表达升高;Sanger Box网站分析结果显示TXNDC12与结直肠癌患者免疫浸润丰度呈负相关(P<0.001)。Western blot及RT-qPCR结果显示,TXNDC12在HT-29、RKO和HCT116细胞中高表达(P<0.05);且敲低TXNDC12后,相较于NC组,si-TXNDC12-2组HT-29和RKO细胞增殖率和克隆形成数量显著下降,细胞迁移距离减少(P<0.05),细胞凋亡增加(P<0.05)。结论TXNDC12在结直肠癌中高表达,且与结直肠癌患者免疫浸润呈负相关,敲低TXNDC12可抑制结直肠癌细胞增殖、迁移并促其凋亡。
Objective To explore the expression of TXNDC12 in colorectal cancer and its effect on proliferation,migration and apoptosis of colorectal cancer cells.Methods The expression of TXNDC12 in colorectal cancer and in colorectal cancer patients with lymph node metastasis were predicted using GEPIA and UALCAN online websites.Eight cases of colorectal cancer tissues and paracancerous tissues from the Affiliated Hospital of Yan'an University were selected to detect the protein expression level of TXNDC12 by Western blot.Sanger Box website was used to analyse the correlation between TXNDC12 expression and the immune infiltration of colon rectal cancer tissues.The protein and mRNA expression levels of TXNDC12 in NCM460,HT-29,RKO and HCT116 cells were detected by Western blot and RT-qPCR.TXNDC12 was knocked down in HT-29 and RKO cells using small interfering RNA(siRNA)technology.The cells were divided into three groups:blank control group(MOCK group),control group(NC group)and knockdown group(si-TXNDC12-2 group),and the proliferation ability,the migration ability and the apoptosis of colorectal cancer cells were detected by CCK-8 assay and clone formation assay,Wound healing assay,Hochest fluorescence staining and Western blot assay,respectively.Results GEPIA analysis and Western blot results of clinical pathological tissues showed that TXNDC12 expression was higher in colo-rectal cancer tissues than that in normal tissues(P<0.05).UALCAN analysis showed that the expression of TXNDC12 was elevated in colorectal cancer patients with lymph node metastasis,and Sanger Box website analysis showed that TXNDC12 was negatively correlated with the abundance of immune infiltration in colorectal cancer patients(P<0.001).Western blot and RT-qPCR results showed that TXNDC12 expression was increased in HT-29,RKO and HCT116 cells compared with NCM460 cells(P<0.05),and compared with NC group,the cell viability,the number of clone formation,and the migration distance of HT-29 and RKO cells in si-TXNDC12-2 group were decreased significantly after knockdown of TXNDC12(P<0.05),while the apoptosis was increased(P<0.05).Conclusion TXNDC12 is highly expressed in colorectal cancer and negatively correlated with immune infiltration in colorectal cancer patients.Knockdown of TXNDC12 can inhibit the cell proliferation and the migration,and promote the apoptosis of colorectal cancer cells.
作者
黑钰
徐向荣
田淑月
陈璇羽
张静
王逢会
HEI Yu;XU Xiangrong;TIAN Shuyue;CHEN Xuanyu;ZHANG Jing;WANG Fenghui(Medical Research and Experimental Center,Yan'an University,Yan'an 716000,China;Yan'an City Fungi Resources Development and Biological Control Key Laboratory)
出处
《山西医科大学学报》
CAS
2024年第1期13-23,共11页
Journal of Shanxi Medical University
基金
陕西省自然科学基础研究计划项目(2021JQ-637)
延安市重点研发项目(2020ZCSY-005)
陕西省高校科协青年人托举计划项目(20200310)
陕西省教育厅突发公共卫生安全专项(20JG041)
陕西省卫生健康科研基金项目(2021D053)。
关键词
结直肠癌
TXNDC12
增殖
迁移
凋亡
colorectal cancer
TXNDC12
proliferation
migration
apoptosis
