【摘要】 目的 探讨KHSRP通过JAK1/STAT3信号通路调节贲门胃癌的恶性发展过程及其分子机制。方法 体外实验选择实时荧光定量聚合酶链式反应(qRT-PCR)来检测胃癌细胞系(MKN-28、OE-33、HGC-27、KYSE-50、CRL-5822、SNU-1)以及人胃黏膜上皮细胞系(GES-1)中KHSRP的表达量。细胞计数试剂盒-8(CCK-8)、Transwell实验分别用于检测细胞增殖、迁移和侵袭情况。裸鼠实验分析KHSRP在活体动物体内的作用。细胞实验和动物实验均分为4组，分别是sh-KHSRP组(KHSRP敲降组),sh-NC组(敲低对照组),KHSRP组(KHSRP过表达组),Vector组(过表达对照组)。Western blot法检测稳定转染细胞JAK/STAT、KHSRP蛋白的表达水平，实时荧光定量PCR测定不同实验组中KHSRP的mRNA表达水平。结果 qRT-PCR显示，与人正常上皮细胞(GES-1)和组织相比，KHSRP在胃癌细胞系(MKN-28、OE-33、HGC-27、KYSE-50、CRL-5822、SNU-1)的表达显著升高。在高表达KHSRP的OE-33、HGC-27细胞系中，敲低KHSRP(sh-KHSRP组)能够显著抑制细胞的增殖、迁移和侵袭能力，而对照组(sh-NC组)无明显抑制效果。在裸鼠实验中，与对照组(sh-NC组)相比，敲低KHSRP表达胃癌细胞的裸鼠显示出肿瘤体积和重量的显著减少，以及肺部转移结节的数量明显减少；接种过表达KHSRP的胃癌细胞则产生相反作用。信号通路实验显示，与sh-NC组相比，sh-KHSRP组JAK1、STAT3的表达量显著下调，过表达组呈相反趋势。细胞拯救实验结果表明KHSRP通过调节JAK1/STAT3信号通路促进胃癌细胞的增殖、迁移和侵袭能力。结论 KHSRP通过调节JAK1/STAT3信号轴，能够促进贲门胃癌转移的恶性进程。更多还原

【Abstract】 Objective To investigate how KHSRP regulates the malignant development process of cardia gastric cancer through JAK1/STAT3 signaling pathway, and to reveal the molecular mechanisms.Methods Quantitative real-time fluorescence polymerase chain reaction(qRT-PCR)was used to detect KHSRP in cancer cell lines(MKN-28,OE-33,HGC-27,KYSE-50,CRL-5822,SNU-1)as well as human normal epithelial cell lines(GES-1).Cell counting kit-8(CCK-8)and Transwell assay were used to detect the biological characteristics of cell proliferation, migration and invasion, respectively.Experimental subjects were divided into four groups in cellular and animal experiments: Sh-KHSRP group(KHSRP knockdown group),Sh-NC group(KHSRP control group),KHSRP group(KHSRP overexpression group),Vector group(KHSRP control group).The expression levels of JAK/STAT and KHSRP proteins in stably transfected cells were detected by Western blotting.The mRNA and protein expression levels of KHSRP in different experimental groups were determined by real-time fluorescence quantitative PCR.Nude mouse experiments were performed to analyze the role of KHSRP in living animals.Results The qRT-PCR showed that the expression of KHSRP was significantly elevated in cancer cell lines(MKN-28,OE-33,HGC-27,KYSE-50,CRL-5822,SNU-1)and tumor tissues compared with human normal epithelial cells(GES-1)and tissues.The results of cell function experiments showed that knockdown of OE-33 and HGC-27 cell lines(sh-KHSRP group)significantly inhibited cell proliferation, migration, and invasive ability, whereas the control group(sh-NC group)did not exhibit this inhibitory effect.These features were significantly enhanced in KHSRP overexpressing cells.In nude mice experiments, knockdown of KHSRP in nude mice led to reduction in tumor volume and weight, lower cell proliferation rate, and lower number of lung nodal metastases compared with the control group(sh-NC group).These changes were reversed while KHSRP was overexpressed.Signaling pathway experiments showed that the expression of JAK1 and STAT3 was significantly downregulated in the sh-KHSRP group compared to the sh-NC group, with opposite results in the overexpression group.The results of cell rescue assay indicated that KHSRP promoted the proliferation, migration and invasion ability of cardia gastric cancer cells by regulating the JAK1/STAT3 signaling pathway.Conclusion KHSRP promotes the malignant process of cardia gastric cancer metastasis by regulating the JAK1/STAT3 signaling axis.更多还原