CAJ | 학술논문

目的构建SET-及MYND-结构域含有蛋白3(SMYD3)荧光质粒并观察组蛋白甲基转移酶SMYD3对肝门部胆管癌细胞生物学行为的影响.方法在人源组织中调取并扩增SMYD3基因酶切并连接于pEGFP-C3质粒构建SMYD3荧光质粒pEGFP-C3-SMYD3.利用质粒转染人肝门部胆管癌细胞株FRH0201并检测SMYD3表达改变,利用细胞计数试剂盒(CCK-8)方法及Transwell方法检测过表达SMYD3后细胞生长及迁移能力变化.结果酶切及测序结果显示SMYD3成功连接入pEGFP-C3载体中,Western blot法结果显示在质粒转染组中SMYD3表达较未转染组升高(73.23±5.60)％,较未转染组及空质粒转染组SMYD3表达水平差异有统计学意义(P＜0.05)；CCK-8方法及Transwell方法显示转染组细胞生长速度较未转染组及空白组加快,迁移能力增强,差异有统计学意义(P＜0.05).结论 SMYD3的过表达促进肝门部胆管癌细胞的增殖及迁移,可能是调控肝门部胆管癌发生发展的因素之一.
목적 구건SET-급MYND-결구역함유단백3(SMYD3)형광질립병관찰조단백갑기전이매SMYD3대간문부담관암세포생물학행위적영향.방법 재인원조직중조취병확증SMYD3기인매절병련접우pEGFP-C3질립구건SMYD3형광질립pEGFP-C3-SMYD3.이용질립전염인간문부담관암세포주FRH0201병검측SMYD3표체개변,이용세포계수시제합(CCK-8)방법급Transwell방법검측과표체SMYD3후세포생장급천이능력변화.결과 매절급측서결과현시SMYD3성공련접입pEGFP-C3재체중,Western blot법결과현시재질립전염조중SMYD3표체교미전염조승고(73.23±5.60)％,교미전염조급공질립전염조SMYD3표체수평차이유통계학의의(P＜0.05)；CCK-8방법급Transwell방법현시전염조세포생장속도교미전염조급공백조가쾌,천이능력증강,차이유통계학의의(P＜0.05).결론 SMYD3적과표체촉진간문부담관암세포적증식급천이,가능시조공간문부담관암발생발전적인소지일.
Objective To construct SET and MYND domainrcontaining protein 3 (SMYD3) fluorescent plasmid and observe the relationship of SMYD3 and biological behaviors of hilar cholangiocarcinoma cells to provide evidence of epigenetic regulation in hilar cholangiocarcinoma.Methods SMYD3 gene was amplified by using polymerase chain reaction (PCR) from human tissue and connected in the plasmid of pEGFP-C3.The hilar cholangiocarcinoma cells were transfected with the plasmid and the expression level of SMYD3 was verified by Western blotting.The growth curve of FRH0201 was examined by cell counting Kit-8 (CCK-8) method and the migratory ability of FRH0201 cells by Transwell.Results The sequencing results showed SMYD3 sequence was successfully connected in the plasmid of pEGFP-C3.Western blotting results showed the expression of SMYD3 was upregulated by (73.23 ± 5.60) ％ in the plasmid transfected group as compared with the empty group and empty plasmid transfected group ( P ＜ 0.05 ).CCK-8 and Transwell showed the growth of cells and the migratory ability were significantly increased in the SMYD3 transfected group as compared with the empty group and empty plasmid transfected group ( P ＜ 0.05 ).Conclusion Over-expression of SMYD3 could increase the proliferation and migratory ability of hilar cholangiocarcinoma cells,which may be one of the important factors in the pathogenesis of hilar cholangiocarcinoma.