钾通道四聚化结构域5对胰腺癌细胞恶性生物学行为的研究

Effects of potassium channel tetramerisation domain containing 5 on the malignant biological behavior of pancreatic cancer cells

目的探讨钾通道四聚化结构域5(KCTD5)对胰腺癌细胞的生物学行为的影响。方法生物信息学检测KCTD5在胰腺癌中184例mRNA及211例蛋白质中的表达水平,观察KCTD5的表达对胰腺癌患者预后的影响,进一步对数据库中所得数据进行单因素Cox回归分析。定量聚合酶链反应(qPCR)、蛋白免疫印迹法检测KCTD5在胰腺癌细胞株中的表达水平。转染KCTD5小干扰RNA至SW1990胰腺癌细胞中,分为si-NC、si-KCTD5-1、si-KCTD5-2组。细胞计数实验(CCK-8)实验检测细胞增殖能力;Transwell实验检测转染后胰腺癌细胞迁移及侵袭能力。结果生物信息学结果表明KCTD5在胰腺癌中的mRNA及蛋白表达水平显著高于正常组织;单因素分析及生存分析结果表明KCTD5可作为胰腺癌患者一种独立危险因素;qPCR结果表明KCTD5在正常人胰腺导管上皮细胞HPDE中的表达水平为1.10±0.18,在胰腺癌细胞株中的表达水平分别为SW1990(2.44±0.19)、PANC-1(1.65±0.15)、ASPC-1(2.15±0.12),KCTD5在胰腺癌细胞株中的表达水平明显升高(t=12.84,5.76,12.16,P均<0.05);转染后,CCK-8实验表明72 h后si-KCTD5-1、si-KCTD5-2、si-KCTD5-3组吸光度值分别为2.93±0.88、1.76±0.97、3.03±0.99,明显低于对照组(4.20±1.02,t=2.31、4.25、2.02,P均<0.05);降低KCTD5表达后Transwell实验中胰腺癌细胞的迁移能力和侵袭能力明显降低。结论胰腺癌中表达升高的KCTD5可促进胰腺癌细胞的增殖、迁移和侵袭能力,降低其表达后胰腺癌细胞上述能力降低。

Objective To investigate the effect of potassium channel tetramerisation domain containing 5(KCTD5)on the biological behavior of pancreatic cancer cells.Methods Bioinformatics was used to analyze the expression level of KCTD5 in 184 mRNAs and 211 proteins in pancreatic cancer and to observe the effect of KCTD5 expression on the prognosis of pancreatic cancer patients,and further to perform one-way Cox regression analysis on the data obtained from the database.Quantitative polymerase chain reaction(qPCR)and protein immunoblotting were used to detect the expression levels of KCTD5 in pancreatic cancer cell lines.KCTD5 small interfering RNA was transfected into SW1990 pancreatic cancer cells and divided into si-NC group,si-KCTD5-1 group and si-KCTD5-2 group.Cell counting kit-8(CCK-8)assay was performed to detect the proliferation ability of cells;the Transwell assay was performed to detect the migration and invasion ability of pancreatic cancer cells after transfection.Results Bioinformatics showed that the mRNA and protein expression levels of KCTD5 were significantly higher in pancreatic cancer than in normal tissues;one-way analysis and survival analysis showed that KCTD5 could be an independent risk factor for pancreatic cancer patients.The expression levels of KCTD5 in normal human pancreatic ductal epithelial cells were(1.10±0.18)and in pancreatic cancer cell lines were SW1990(2.44±0.19),PANC-1(1.65±0.15)and ASPC-1(2.15±0.12),respectively.qPCR results showed that the expression levels of KCTD5 in pancreatic cancer cell lines were(1.10±0.18),and the expression levels of KCTD5 in pancreatic cancer cell lines were(2.44±0.19),PANC-1(1.65±0.15)and ASPC-1(2.15±0.12),respectively.cell lines showed significantly higher expression levels(t=12.84,5.76,12.16,all P<0.05);after transfection,CCK-8 experiments showed that the absorbance values after 72 h were(4.20±1.02)in the si-NC group,(2.93±0.88)in the si-KCTD5-1 group,(1.76±0.97)in the si-KCTD5-2 group,(1.76±0.97)in the si-KCTD5-3 group(3.03±0.99),si-KCTD5-1 group,si-KCTD5-2 group and si-KCTD5-3 group were significantly lower(t=2.31,4.25,2.02,all P<0.05)compared with the control group(4.20±1.02);the pancreatic cancer cells in Transwell assay after reducing KCTD5 expression The migration ability and invasion ability of pancreatic cancer cells in Transwell assay were significantly reduced after reducing KCTD5 expression.Conclusion Elevated expression of KCTD5 in pancreatic cancer promotes the proliferation,migration and invasion ability of pancreatic cancer cells,and the above-mentioned ability of pancreatic cancer cells is reduced after decreasing its expression.