FAM3C表达对胰腺癌细胞增殖、迁移、侵袭的影响和调控机制

Effects and regulatory mechanisms of FAM3C expression on proliferation,migration,and invasion of pancreatic cancer cells

目的分析序列相似家族3成员C(FAM3C)在胰腺癌中的表达以及对胰腺癌细胞增殖、迁移、侵袭等影响和相关机制。方法收集2022年7月至2023年6月在江南大学附属医院手术切除的4例胰腺癌患者的癌组织和癌旁组织。聚合酶链反应和Western印迹检测组织和胰腺癌细胞PANC-1、MIA PaCa-2中FAM3C的表达。选取PANC-1、MIA PaCa-2细胞(FAM3C高表达)设置FAM3C-siRNA干扰组和甲基转移酶样蛋白3(METTL3)-siRNA干扰组,予以相应干扰小RNA处理,同时设置阴性对照组。细胞计数检测增殖能力,Transwell检测细胞迁移及侵袭。免疫共沉淀甲基化抗体检测FAM3C是否发生甲基化以及METTL3是否介导此甲基化。结果4例胰腺癌患者癌组织中FAM3C的mRNA相对表达为(1.29±0.19),高于癌旁组织(0.47±0.17),差异有统计学意义(t=-6.48,P=0.001)。与阴性对照A组相比,FAM3C-siRNA干扰组增殖能力下降,迁移和侵袭细胞数减少,差异均有统计学意义(均P<0.05)。在PANC-1细胞阴性对照B组中,IgG处理的细胞FAM3C mRNA相对表达为(1.05±0.53),低于甲基化抗体处理组的(30.57±1.09),差异有统计学意义(t=-42.04,P=0.001)。PANC-1细胞经甲基化抗体处理后,阴性对照B组FAM3C mRNA相对表达高于METTL3-siRNA干扰组(30.57±1.09)比(18.17±0.50),差异有统计学意义(t=17.89,P=0.001)。与阴性对照B组相比,METTL3-siRNA干扰组PANC-1细胞增殖能力、迁移和侵袭细胞数降低,差异均有统计学意义(均P<0.05)。MIA PaCa-2与PANC-1细胞检测结果一致。结论FAM3C在胰腺癌中高表达,且促进胰腺癌细胞增殖、迁移和侵袭,METTL3通过甲基化修饰FAM3C影响其表达,进一步影响胰腺癌细胞增殖、迁移和侵袭。

ObjectiveTo analyze the expression of sequence similarity family 3 member C(FAM3C)in pancreatic cancer and its effects on pancreatic cancer cell proliferation,migration,invasion and the underlying mechanisms.MethodsTissue samples from four pancreatic cancer patients undergoing surgical resection at Affiliated Hospital of Jiangnan University from July 2022 to June 2023 were collected.Expression of FAM3C was assessed in both cancer and adjacent tissues using polymerase chain reaction and Western blot techniques.High FAM3C-expressing pancreatic cancer cells PANC-1 and MIA PaCa-2 were selected for further experiments involving FAM3C-siRNA and methyltransferase-like protein 3(METTL3)-siRNA groups,alongside a negative control group.Cell counting,Transwell assays,and co-immunoprecipitation with methylation antibodies were utilized to evaluate the proliferation,migration,invasion,and methylation status of FAM3C.ResultsThe relative expression of FAM3C mRNA in cancer tissues of the four patients with pancreatic cancer was higher than that in adjacent tissues(1.29±0.19 vs 0.47±0.17,t=-6.48,P=0.001).Compared with the negative control group A,the proliferation ability of the FAM3C-siRNA interference group decreased,and the number of migrating and invasive cells decreased(all P<0.05).In the negative control group B of PANC-1 cells,the relative expression of FAM3C mRNA in IgG-treated cells was lower than that in the methylated antibody-treated group(1.05±0.53 vs 30.57±1.09,t=-42.04,P=0.001).After PANC-1 cells were treated with methylated antibodies,the relative expression of FAM3C mRNA in the negative control group B was higher than that in the METTL3-siRNA interference group(30.57±1.09 vs 18.17±0.50,t=17.89,P=0.001).Compared with the negative control group B,the proliferation ability,migration and invasion cell numbers of PANC-1 cells in the METTL3-siRNA interference group were reduced(all P<0.05).The results of MIA PaCa-2 and PANC-1 cells were consistent.ConclusionsFAM3C is highly expressed in pancreatic cancer and promotes the proliferation,migration and invasion of pancreatic cancer cells.METTL3 affects FAM3C expression by methylation and proliferation,migration and invasion of pancreatic cancer cells.