谷氨酰胺酶1介导主穹隆蛋白表达调控肝内胆管癌细胞对5-氟尿嘧啶化疗敏感性的研究

Glutamine 1 regulates the chemosensitivity of intrahepatic cholangiocarcinoma cells to 5-FU by targeting major vault protein

目的探讨谷氨酰胺酶1(glutamine1,GLS1)对肝内胆管癌细胞5-氟尿嘧啶化疗敏感性的影响及其机制(方法生物信息数据库检索胆管癌中GLS1和主穹隆蛋白(major vault protein,MVP)表达水平及两者的相关性。Western blot法、免疫组化检测42例肝内胆管细胞癌组织GLS1和MVP表达水平,并统计两者相关性;运用siRNA干扰及pcDNA过表达调控胆管癌细胞株GLS1表达,Western blot验证干扰及过表达效率及MVP表达;最后使用细胞计数试剂盒(CCK-8)法检测各组细胞对化疗药物5-氟尿嘧啶的化疗敏感性.结果在胆管癌组织中GLS1和MVP表达显著上调(tCLSI=3.963;tmvp=3.131,P<0.05),并且GLS1表达与MVP呈正相关(r^2=0.3517,P<0.05)。干扰QBC939细胞GLS1表达后可下调MVP表达,并显著提升细胞对5-氟尿嘧啶化疗敏感性。反之,过表达RBE细胞GLS1表达可上调MVP表达,并降低细胞对5-氟尿嘧啶化疗敏感性。结论GLS1可调控胆管癌细胞对5-氟尿嘧啶化疗敏感性,其机制可能与其调控MVP表达有关。

Objective To study the functions and mechanisms of glutamine 1 ( GLS1 ) in intrahepatic cholangiocarcinoma ( ICC ) cell to 5-fluorouraeil ( 5-FU ) chemosensitivity. Methods The expression and relation between GLS1 and major vault protein ( MVP) in cholangiocarcinoma were analyzed by bioinformatics database. Western blot and immunohistochemistry were used to detect the expression of GLS1 and MVP in 42 ICC tissues, and the correlation between GLS1 and MVP was studied by statistics. The regulation of GLS1 in ICC cell were evaluated by siRNA interference and pcDNA overexpression, and then tested the interference and overexpression efficiency of GLS1 by Western blotting. The chemosensitivity to 5-Fu was tested by cell counting kit-8( CCK-8). Results The expression of GLS1 and MVP in ICC tissues was significantly up-regulated (tGLS1 = 3. 963;tMVP=3. 131, P < 0. 05 ), and the expression of GLS1 was positively correlated with MVP( r^2 = 0. 351 7 , P < 0. 05 ). Knockdown of GLS1 in QBC939 cells enhanced chemosensitivity of QBC939 cells to 5-Fu and notably downregulated MVP expression, while enforced expression of GLS1 in RBE cells promoted MVP expression and reduce cell sensitivity to 5-fluorouracil chemosensitivity. Conclusions GLS1 regulates the chemosensitivity of ICC cells to 5-Fu, and its mechanism may relates to the regulation of MVP.