KH-type splicing regulatory protein (KHSRP) plays an important role in cancer invasion, but the relevant mechanism is not well known. In the present study, we investigated the function and potential molecular mechanism of KHSRP in non-small cell lung cancer (NSCLC) metastasis and elucidated its clinical significance. Isobaric tags for relative and absolute quantitation and the SWATH™ approach were combined with nanoliquid chromatography-tandem mass spectrometry analysis to identify metastasis-associated nucleoproteins in NSCLC. Real-time PCR and Western blot were used to screen for metastasis-associated candidate molecules. Gene knockdown and overexpression were used to investigate their functions and molecular mechanisms in lung cancer cells. Coimmunoprecipitation (Co-IP) experiments were performed to identify the interactions between candidate molecules and their interacting proteins. Gene expression and its association with multiple clinicopathologic characteristics were analyzed by immunohistochemistry (IHC) and Western blot in human lung cancer specimens. KHSRP was identified as a metastasis-associated candidate molecule. In NSCLC cell lines, knockdown of KHSRP significantly reduced lung cancer cell proliferation, migration, and invasion in vitro and in vivo, whereas overexpression of KHSRP did the opposite. Mechanistically, the protein heterogeneous nuclear ribonucleoprotein C (C1/C2) (HNRNPC) was identified to interact with KHSRP using Co-IP experiments. In NSCLC cell lines, overexpression of HNRNPC significantly promoted lung cancer cell proliferation, migration, and invasion in vitro and in vivo. KHSRP and HNRNPC may induce human lung cancer cell invasion and metastasis by activating the IFN-α-JAK-STAT1 signaling pathway. Drastically higher expression levels of KHSRP and HNRNPC were observed in lung cancer tissues compared to those in adjacent noncancerous tissues. Increased KHSRP and HNRNPC expression was significantly associated with advanced tumor stages and metastasis (both lymph node and distant). Kaplan-Meier survival analysis showed that patients with high KHSRP and HNRNPC expression levels were predicted to have the shortest survival times and to have a poor prognosis. KHSRP plays an important role in NSCLC metastasis and may serve as a potential prognostic marker and novel therapeutic target for lung cancer metastasis treatment.

中文翻译：

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RNA结合蛋白KHSRP促进非小细胞肺癌的肿瘤生长和转移。

KH型剪接调节蛋白（KHSRP）在癌症侵袭中起着重要作用，但相关机制尚不清楚。在本研究中，我们研究了KHSRP在非小细胞肺癌（NSCLC）转移中的功能和潜在的分子机制，并阐明了其临床意义。将用于相对定量和绝对定量的等压标签和SWATH™方法与纳米液相色谱-串联质谱分析相结合，以鉴定NSCLC中与转移相关的核蛋白。实时PCR和蛋白质印迹用于筛选与转移相关的候选分子。基因敲低和过表达被用来研究其功能和分子机制在肺癌细胞中。进行了共免疫沉淀（Co-IP）实验，以鉴定候选分子与其相互作用蛋白之间的相互作用。通过免疫组化（IHC）和蛋白质印迹分析了人类肺癌标本中的基因表达及其与多种临床病理特征的关系。KHSRP被确定为与转移相关的候选分子。在NSCLC细胞系中，KHSRP的敲低显着降低了肺癌细胞的体外，体内增殖，迁移和侵袭，而KHSRP的过表达则相反。从机理上讲，使用Co-IP实验确定了蛋白质异质核糖核蛋白C（C1 / C2）（HNRNPC）与KHSRP相互作用。在NSCLC细胞系中，HNRNPC的过度表达显着促进了肺癌细胞的增殖，迁移，和体外和体内入侵。KHSRP和HNRNPC可能通过激活IFN-α-JAK-STAT1信号通路诱导人肺癌细胞侵袭和转移。与邻近的非癌组织相比，在肺癌组织中观察到KHSRP和HNRNPC的表达水平大大提高。KHSRP和HNRNPC表达增加与晚期肿瘤分期和转移（淋巴结转移和远处转移）显着相关。Kaplan-Meier生存分析表明，具有高KHSRP和HNRNPC表达水平的患者预计生存时间最短，预后也较差。KHSRP在NSCLC转移中起着重要作用，并可能成为肺癌转移的潜在预后标志物和新的治疗靶标。KHSRP和HNRNPC可能通过激活IFN-α-JAK-STAT1信号通路诱导人肺癌细胞侵袭和转移。与邻近的非癌组织相比，在肺癌组织中观察到KHSRP和HNRNPC的表达水平大大提高。KHSRP和HNRNPC表达增加与晚期肿瘤分期和转移（淋巴结转移和远处转移）显着相关。Kaplan-Meier生存分析表明，具有高KHSRP和HNRNPC表达水平的患者预计生存时间最短，预后也较差。KHSRP在NSCLC转移中起着重要作用，并可能成为肺癌转移的潜在预后标志物和新的治疗靶标。KHSRP和HNRNPC可能通过激活IFN-α-JAK-STAT1信号通路诱导人肺癌细胞侵袭和转移。与邻近的非癌组织相比，在肺癌组织中观察到KHSRP和HNRNPC的表达水平大大提高。KHSRP和HNRNPC表达增加与晚期肿瘤分期和转移（淋巴结转移和远处转移）显着相关。Kaplan-Meier生存分析表明，具有高KHSRP和HNRNPC表达水平的患者预计生存时间最短，预后也较差。KHSRP在NSCLC转移中起着重要作用，并可能成为肺癌转移的潜在预后标志物和新的治疗靶标。与邻近的非癌组织相比，在肺癌组织中观察到KHSRP和HNRNPC的表达水平大大提高。KHSRP和HNRNPC表达增加与晚期肿瘤分期和转移（淋巴结转移和远处转移）显着相关。Kaplan-Meier生存分析表明，具有高KHSRP和HNRNPC表达水平的患者预计生存时间最短，预后也较差。KHSRP在NSCLC转移中起着重要作用，并可能成为肺癌转移的潜在预后标志物和新的治疗靶标。与邻近的非癌组织相比，在肺癌组织中观察到KHSRP和HNRNPC的表达水平大大提高。KHSRP和HNRNPC表达增加与晚期肿瘤分期和转移（淋巴结转移和远处转移）显着相关。Kaplan-Meier生存分析表明，具有高KHSRP和HNRNPC表达水平的患者预计生存时间最短，预后也较差。KHSRP在NSCLC转移中起着重要作用，并可能成为肺癌转移的潜在预后标志物和新的治疗靶标。KHSRP和HNRNPC表达增加与晚期肿瘤分期和转移（淋巴结转移和远处转移）显着相关。Kaplan-Meier生存分析表明，具有高KHSRP和HNRNPC表达水平的患者预计生存时间最短，预后也较差。KHSRP在NSCLC转移中起着重要作用，并且可以作为潜在的预后标志物和肺癌转移治疗的新治疗靶标。KHSRP和HNRNPC表达增加与晚期肿瘤分期和转移（淋巴结转移和远处转移）显着相关。Kaplan-Meier生存分析表明，具有高KHSRP和HNRNPC表达水平的患者预计生存时间最短，预后也较差。KHSRP在NSCLC转移中起着重要作用，并可能成为肺癌转移的潜在预后标志物和新的治疗靶标。