CLDN9与TYK2在无功能性垂体腺瘤的表达及与侵袭性的相关性

Expressions of CLDN9 and TYK2 in non-functioning pituitary adenoma correlated with invasiveness

目的 研究Claudin9(CLDN9)、酪氨酸激酶2(tyrosine kinase 2,TYK2)在无功能性垂体腺瘤(non-functioning pituitary adenoma,NFPA)中的表达与侵袭性的相关性,以及CLDN9与TYK2在侵袭性NFPA中的相互作用。方法 收集2020-06—2021-05在新疆维吾尔自治区人民医院手术治疗的24例NFPA患者的临床资料及手术标本,根据术前影像检查及术中所见信息分为侵袭性NFPA组和非侵袭性NFPA组各12例。通过实时定量聚合酶链反应(quantitative real-time polymerase chain reaction,qRT-PCR)、免疫组化(immunohistochemistry,IHC)及蛋白质印迹(Western blot,WB)等方法检测CLDN9、TYK2在2组NFPA组织中的表达量;用慢病毒质粒转染技术构建垂体瘤GT1-1细胞系空载对照组(NC组)和CLDN9过表达实验组(OE-CLDN9组),用WB和qRT-PCR验证CLDN9对垂体瘤GT1-1细胞系的稳定转染。通过qRT-PCR检测TYK2在NC和OE-CLDN9等2组细胞中的表达量。结果 qRT-PCR、IHC及WB结果均显示,CLDN9在侵袭性NFPA组织中的表达量明显高于非侵袭性NFPA,差异均有统计学意义(t=12.63、17.98、2.45,P<0.001),而TYK2在侵袭性NFPA组织中的表达量明显低于非侵袭性NFPA,差异均有统计学意义(t=13.82、4.29、2.26,P<0.001)。检测结果显示,OE-CLDN9组中的CLDN9表达量明显高于NC组(F=943.30,P<0.001),TYK2表达量明显低于NC组(F=13.64,P=0.004)。结论 CLDN9表达上调与NFPA侵袭性生长有关,且可能通过低表达的TYK2促进NFPA侵袭性。

Objective To evaluate the correlation among the expression of claudin-9(CLDN9),tyrosine kinase-2(TYK2) and non-functioning pituitary adenoma(NFPA) invasiveness,and their associations.Methods Clinical data and surgical specimens of 24 patients with NFPA surgically treated in our hospital from June 2020 to May 2021 were collected and divided into invasive and non-invasive NFPA groups based on their pre-operative imaging and intra-operative information,comprising 12 patients for each group.CLDN9 and TYK2 expressions in the NFPA tissues of the two groups were measured by quantitative real-time polymerase chain reaction(qRT-PCR),immunohistochemistry(IHC) and Western blotting(WB).The lentiviral plasmid transfection technique was used to develop a rat pituitary tumour GT1-1 cell line null control group(NC) and CLDN9 overexpression experimental group(OE-CLDN9),and the stable transfection of CLDN9 to the rat pituitary tumour G1 l-1 cell line was detected using WB and qRT-PCR.TYK2 expressions in the NC and OE-CLDN9 cell groups were detected using qRT-PCR.Results The qRT-PCR,IHC and WB results revealed that the expression of CLDN9 was significantly higher in invasive than in non-invasive NFPA tissues,and the expression differences were statistically significant(t=12.63,17.98,2.45,P<0.001),while the expression of TYK2 was significantly lower in invasive than in non-invasive NFPA tissues,and the expression differences were statistically significant(t=-13.82,4.29,2.26,P<0.001).The detected results showed that CLDN9 expression in the OE-CLDN9 group were significantly higher than in the NC group(F=943.30,P<0.001),TYK2 expression was significantly lower in the OE-CLDN9 group than in the NC group(F=13.64,P=0.004).Conclusion Upregulated CLDN9 expression is associated with NFPA invasive growth and may promote NFPA invasiveness through low expression of TYK2.