硫氧还蛋白结构域包含蛋白9在胆囊癌组织的表达及其对胆囊癌细胞增殖和侵袭的影响

Expression of thioredoxin domain-containing protein 9 in gallbladder cancer and its effect on cell proliferation and invasion

目的:探讨硫氧还蛋白结构域包含蛋白9(TXNDC9)在胆囊癌组织中的表达水平及其对胆囊癌细胞增殖和侵袭的影响。方法:收集2017年1月至2022年12月浙江大学医学院附属第四医院手术切除的48例胆囊癌患者肿瘤组织和癌旁组织标本。通过免疫组织化学染色法检测TXNDC9蛋白表达,分析其与患者临床病理特征的相关性。采用RNA干扰技术沉默胆囊癌GBC-SD细胞中TXNDC9基因表达,细胞计数法(CCK-8)和Transwell实验检测细胞的增殖和侵袭能力,蛋白质印迹法(Western blot)检测磷酸化磷脂酰肌醇3激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(vimentin)的表达水平。组间比较采用χ^(2)检验和独立样本t检验。结果:胆囊癌组织中TXNDC9蛋白的阳性表达率明显高于癌旁组织[70.8%(34/48)比31.3%(15/48),χ^(2)=15.048,P<0.05],且TXNDC9表达与肿瘤分期(χ^(2)=9.708,P<0.05)和淋巴结转移(χ^(2)=7.987,P<0.05)显著相关。CCK-8实验结果表明,TXNDC9敲减组细胞的增殖能力显著低于对照组(吸光度值:0.453±0.054比0.822±0.085,t=8.991,P<0.05)。Transwell实验结果表明,TXNDC9敲减组细胞的侵袭能力显著低于对照组[穿膜细胞数:(71.800±8.526)个比(183.200±16.724)个,t=13.269,P<0.05]。Western blot实验结果表明,TXNDC9敲减组p-PI3K、p-Akt、N-cadherin、vimentin的相对表达量(0.224±0.056、0.238±0.046、0.273±0.037、0.386±0.045)均显著低于对照组(0.812±0.077、0.837±0.092、0.748±0.069、0.807±0.085),差异有统计学意义(t=10.684、10.138、10.471、7.567,P<0.05);TXNDC9敲减组E-cadherin的相对表达量显著高于对照组(0.898±0.095比0.294±0.039,t=10.209,P<0.05)。结论:TXNDC9在胆囊癌中表达升高,与肿瘤进展密切相关,并可通过PI3K/Akt信号通路调控胆囊癌细胞的增殖和侵袭能力。

Objective To investigate the expression of thioredoxin domain-containing protein 9(TXNDC9)in gallbladder cancer(GBC)and its effect on cell proliferation and invasion.Methods The surgically resected specimens of 48 GBC patients were collected in the Fouth Affiliated Hospital of Zhejiang University School of Medicine from January 2017 to December 2022.The expression of TXNDC9 protein in GBC tissues and corresponding paracancer paracancerous tissues was detected by immunohistochemistry.Associations between TXNDC9 expression and clinicopathological features were analyzed.RNA interference technology was employed to silence the expression of TXNDC9 gene in GBC-SD cells.Cell counting kit-8(CCK-8)and Transwell assays were performed to evaluate the cell proliferation and invasion ability abilities.The expressions of phosphorylated PI3K(p-PI3K),phosphorylated Akt(p-Akt),E-cadherin,N-cadherin and vimentin were detected by Western blotting.comparisons between groups were performed by using the chi-squareχ^(2) test and independent sample t-test.Results The positive expression rate of TXNDC9 protein in GBC tissues was significantly higher than that in paracancerous tissues[70.8%(34/48)vs.31.3%(15/48),χ^(2)=15.048,P<0.05].Furthermore,TXNDC9 expression was significantly associated with tumor stage(χ^(2)=9.708,P<0.05)and lymph nodal metastasis(χ^(2)=7.987,P<0.05).CCK-8 assay indicated that the cell proliferation ability in TXNDC9 knockdown group significantly decreased than that in the control group(absorbance value:0.453±0.054 vs.0.822±0.085,t=8.991,P<0.05).Transwell assay showed that the cell invasion ability in TXNDC9 knockdown group significantly decreased than that in the control group[number of transmembrane cells:(71.800±8.526)cells vs.(183.200±16.724)cells,t=13.269,P<0.05].Western blotting analysis revealed that the relative expressions of p-PI3K,p-Akt,N-cadherin,vimentin(0.224±0.056,0.238±0.046,0.273±0.037,0.386±0.045)in TXNDC9 knockdown group were significantly lower than those(0.812±0.077,0.837±0.092,0.748±0.069,0.807±0.085)in the control group,and the differences were statistically significant(t=10.684,10.138,10.471,7.567,all P<0.05);the relative expression of E-cadherin in TXNDC9 knockdown group was significantly higher than that in the control group(0.898±0.095 vs.0.294±0.039,t=10.209,P<0.05).Conclusion TXNDC9 expression is upregulated in GBC and correlated with tumor progression.TXNDC9 can promote the proliferation and invasion of GBC cells through the PI3K/Akt signaling pathway.