目的 探讨LINC00626靶向调控JAK1/STAT3/KHSRP轴在胃癌恶性生物学行为中的潜在作用机制。 方法 采用qRT-PCR 检测胃癌细胞系（AGS、MGC-803、NCI-87、BGC-823、MNK-45、SGC-7901）以及人正常胃癌上皮细胞系（GES-1）和120例胃癌组织标本及其正常组织标本中LINC00626、KHSRPmRNA的表达量。细胞计数试剂盒-8（CCK-8）、划痕和Transwell迁移/侵袭实验分别检测细胞增殖、迁移和侵袭等生物学特征。采用Western blot法检测稳定转染细胞中JAK/STAT、KHSRP蛋白的表达水平。裸鼠实验分析LINC00626在活体动物体内的作用。 结果 qRT-PCR显示，与人正常胃癌细胞（GES-1）和组织相比，LINC00626和KHSRP在胃癌细胞系（AGS、MGC-803、NCI-87、BGC-823、MNK-45、SGC-7901）和肿瘤组织中的表达显著升高。细胞功能实验显示，LINC00626过表达会促进胃癌细胞系MNK-45和SGC-7901的增殖、迁移和侵袭，KHSRP能够使敲降LINC00626的胃癌细胞增殖、迁移和侵袭能力得到恢复。体内动物实验结果显示，与对照组相比，sh-LINC00626组裸鼠体内肿瘤的体积和重量、细胞增殖率和胃癌肺转移病灶数目明显减少；过表达结果相反。信号通路实验显示，与sh-NC组相比，sh-LINC00626组中的JAK1、STAT3mRNA的表达水平呈明显下降趋势，过表达组结果相反；与sh-NC组相比，sh-KHSRP组JAK1、STAT3的表达量显著下调，过表达组结果相反。 结论 LINC00626由JAK1/STAT3/KHSRP信号传导通路，加速了胃癌的恶性转移进程。
Abstract
Objective To investigate the potential mechanism of LINC00626-targeted regulation of the JAK1/STAT3/KHSRP axis in the malignant biological behavior of gastric cancer. Methods qRT-PCR was used to detect the expression levels of LINC00626 and KHSRP mRNA in gastric cancer cell lines (AGS, MGC-803, NCI-87, BGC-823, MNK-45, SGC-7901) and human normal gastric epithelial cell line (GES-1) and 120 cases of gastric cancer tissue specimens and normal tissue specimens. The biological characteristics of cell proliferation, migration and invasion were detected by cell counting kit-8 (CCK-8), scratch and Transwell migration/invasion assays, respectively. The expression levels of JAK/STAT and KHSRP proteins in stably transfected cells were detected by Western blot. The role of LINC00626 in vivo was analyzed in nude mice. Results qRT-PCR results showed that the expression of LINC00626 and KHSRP in gastric cancer cell lines (AGS, MGC-803, NCI-87, BGC-823, MNK-45, SGC-7901) and tumor tissues was significantly higher than that in human normal gastric cancer cells (GES-1) and tissues. Cell function experiments showed that LINC00626 overexpression promoted the proliferation, migration and invasion of gastric cancer cell lines MNK-45 and SGC-7901, and KHSRP was able to restore the ability of proliferation, migration and invasion in gastric cancer cells with knockdown of LINC00626. The results of in vivo animal experiments showed that, compared with the sh-NC group, the tumor volume and weight, cell proliferation rate and the number of lung metastases of gastric cancer in the sh-LINC00626 group were significantly reduced, while the overexpression results were opposite. Signaling pathway experiments showed that compared with the sh-NC group, the expression levels of JAK1 and STAT3 mRNA in the sh-LINC00626 group showed a significant downward trend, while the results in the overexpression group were opposite. Compared with the sh-NC group, the expression of JAK1 and STAT3 in the sh-KHSRP group was significantly down-regulated, and the results in the overexpression group were opposite. Conclusions LINC00626 accelerates the malignant metastasis of gastric cancer by JAK1/STAT3/KHSRP signaling pathway.
关键词
胃癌； /
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LINC00626； /
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JAK1； /
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STAT3； /
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KHSRP
Key words
GC; /
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LINC00626; /
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JAK1; /
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STAT3; /
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KHSRP
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基金
河南省教育厅项目（25B320018）；河南省科技厅科技攻关项目（252102311055、252102311053）
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