基因相似序列家族成员126A靶向调控波形蛋白促进胰腺癌细胞侵袭和迁移及其机制

Gene similar sequence family member 126A targets vimentin to promote invasion and migration of pancreatic cancer cells and its mechanism

目的:探讨基因相似序列家族成员126A(FAM126A)靶向波形蛋白(Vimentin)对胰腺癌侵袭和迁移能力的影响。方法:利用基因表达谱交互分析数据库(GEPIA2)和泛癌长期预后和基因表达谱数据库(LOGpc)中的数据,分析FAM126A在胰腺癌和癌旁组织中的表达差异以及对胰腺癌患者生存期的影响;选取2020年1月至2022年1月收集的胰腺导管癌肿瘤组织和对应的癌旁组织样本共16对作为研究对象,采用蛋白印迹法(Western blot)分析胰腺癌组织和癌旁组织中FAM126A表达水平;在胰腺癌PANC-1细胞系构建2个小干扰RNA(siRNA)序列并建立si-FAM126A#1、si-FAM126A#2沉默组和si-con对照组,采用细胞划痕和Transwell实验检测沉默FAM126A后对胰腺癌细胞侵袭和迁移能力的影响;Western blot检测上皮-间充质转化(EMT)相关蛋白E-钙黏蛋白(E-candberin)、波形蛋白(Vimentin)和Snail蛋白的表达量;采用生物信息学和蛋白质谱检测报告分析FAM126A的靶基因,并通过共定位和共沉淀的方法进一步证实FAM126A与Vimentin的相互作用关系。计量数据资料采用均值±标准差(Mean±SD)进行描述,组间比较采用t检验。P<0.05为差异有统计学意义。结果:在胰腺癌组织中FAM126A表达明显高于癌旁组织(t=12.880,P<0.05);沉默组si-FAM126A#1、si-FAM126A#2胰腺癌细胞侵袭细胞数(66.11±8.68,91.11±8.68)和迁移率[(22.56±5.36)%,(39.11±6.91)%]较对照组si-con[(146.00±9.15)%,(75.01±10.36)%]显著降低(P<0.05);si-FAM126A#1和si-FAM126A#2组E-candberin蛋白相对表达量(1.20±0.06,0.97±0.14)较si-con组(0.71±0.08)显著升高(P<0.05),而Vimentin、Snail蛋白表达量为(0.92±0.09,1.01±0.15;0.49±0.07,0.55±0.08)均显著高于对照组(1.68±0.11;1.21±0.08)(P<0.05);生物信息学和蛋白质谱检测报告提示FAM126A与Vimentin存在相互作用关系,共定位和免疫共沉淀证实了Vimentin是FAM126A的靶蛋白。结论:在胰腺癌组织中FAM126A的表达高于癌旁组织,而FAM126A可通过靶向调控Vimentin的表达诱导胰腺癌细胞EMT进程,最终促进胰腺癌细胞的侵袭和迁移能力。

Objective To investigate the effects of 126A(FAM126A)targeting Vimentin on invasion and migration of pancreatic cancer Methods The expression difference of FAM126A in pancreatic and paracancer tssues and its effect on survival of pancreatic cancer patients were analyzed using data from thegene expression profle Interactive analysis database (GEPIA2) and the pancarcinoma Long-Term Prognosisand Gene expression profile database (LOGpc). A total of 16 pairs of pancreatic ductal carcinoma tumortissues and corresponding paracancer tissue samples collected from January 2020 to January 2022 wereselected as research objects. Western blot was used to analyze the expression level of FAM126A in pancreaticand paracancer tissues. Two small interfering RNA (siRNA) sequences were constructed in PANC-1 cell lineof pancreatic cancer and si-FAM126A#1, si-FAM126A#2 silencing groups and si-con control group wereestablished. The effects of silencing FAM126A on the invasion and migration of pancreatic cancer cells weredetected by cell scratch and Transwell assay. The expression levels of EMT-related proteins E-candberin,Vimentin and Snail were detected by Western blot. The target gene of FAM126A was analyzed bybioinformatics and protein spectrogram reports, and the interaction between FAM126A and Vimentin wasfurther confirmed by co-localization and co-precipitation methods. Measurement data were described by MeantSD, and t test was used for comparison between groups. Results The expression of FAM126A in pancreaticcancer tissues was significantly higher than that in adjacent tissues (t=12. 880, P<0. 05). The number ofpancreatic cancer cells invaded by si-FAM126A#1 and si-FAM126A#2 groups (66. 11+8. 68, 91. 11±8. 68)and mobility [(22. 56±5.36) %, (39.11±6.91) %] was significantly lower than that of control group si-con[(146. 00±9. 15) %, (75. 01±10. 36) %] (P<0. 05). The relative expression level of E-candberin protein insi-FAM126A#1 and si-FAM126A#2 groups was significantly increased (1.20±0.06, 0. 97±0.14) comparedwith that in si-con group (0.71±0. 08) (P<0.05). Vimentin Snail protein expression levels were (0. 92±0.09, 1.01±0.15;0. 49±0.07, 0.55+0.08) were significantly higher than the control group (1.68±0. 11;1.21+0.08) (P<0.05);Bioinformatics and protein spectrogram reports suggested that there was aninteraction between FAM126A and Vimentin. Co-localization and co-immunoprecipitation confirmed thatVimentin was the target protein of FAM126A. Conclusion The expression of FAM126A in pancreatic cancertissues is higher than that in adjacent tissues, and FAM126A can induce the EMT process of pancreatic cancercells through targeted regulation of Vimentin expression, and finally promote the invasion and migration abilityof pancreatic cancer cells.