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Abstract：
BACKGROUND: CLCA2 was reported as a tumor suppressor and disregulated in breast cancer. However, its function in tumor growth and metastasis in NPC has rarely been reported. In this study, we investigated the functional and molecular mechanisms by which CLCA2 influences NPC.
METHODS: CLCA2 expression in human NPC cell lines and tissues was examined via real-time PCR (RT-PCR), Western blot and IHC. The biological roles of CLCA2 in proliferative, migration and invasion of NPC cell lines was evaluated in 5-8F, S18, S26 and SUNE-1 cells. Cell viability, migration and invasion were assessed in vitro by MTS, colony formation and transwell assay, respectively. CLCA2 in growth and metastasis of NPC were evaluated in vivo through NPC xenograft tumor growth, lung metastatic mice model and popliteal lymph node (LN) metastasis model.
RESULTS: Overexpression of CLCA2 significantly decreased proliferation, migration and invasion of NPC cells. In contrast, knockdown of CLCA2 elicited the opposite effects. CLCA2 overexpression suppressed xenograft tumor growth and lung, popliteal lymph node (LN) metastasis in vivo. CLCA2 inhibited tumor metastasis through suppressing epithelial-Mesenchymal transition (EMT) and in-activating FAK/ERK1/2 signaling pathway in NPC cells. Immunohistochemical staining of 143 NPC samples revealed that CLCA2 expression was an independent, favorable prognostic factor for overall survival and distant metastasis-free survival of patients. In addition, inhibition of FAK and ERK1/2 reversed CLCA2 silencing-induced tumor cell migration. Furthermore, inhibitors against chloride channels suppressed NPC cellular migration which could have been enhanced by the presence of CLCA2.
CONCLUSIONS: CLCA2 suppress NPC proliferation, migration, invasion and epithelial-mesenchymal transition through inhibiting FAK/ERK signaling.
METHODS: CLCA2 expression in human NPC cell lines and tissues was examined via real-time PCR (RT-PCR), Western blot and IHC. The biological roles of CLCA2 in proliferative, migration and invasion of NPC cell lines was evaluated in 5-8F, S18, S26 and SUNE-1 cells. Cell viability, migration and invasion were assessed in vitro by MTS, colony formation and transwell assay, respectively. CLCA2 in growth and metastasis of NPC were evaluated in vivo through NPC xenograft tumor growth, lung metastatic mice model and popliteal lymph node (LN) metastasis model.
RESULTS: Overexpression of CLCA2 significantly decreased proliferation, migration and invasion of NPC cells. In contrast, knockdown of CLCA2 elicited the opposite effects. CLCA2 overexpression suppressed xenograft tumor growth and lung, popliteal lymph node (LN) metastasis in vivo. CLCA2 inhibited tumor metastasis through suppressing epithelial-Mesenchymal transition (EMT) and in-activating FAK/ERK1/2 signaling pathway in NPC cells. Immunohistochemical staining of 143 NPC samples revealed that CLCA2 expression was an independent, favorable prognostic factor for overall survival and distant metastasis-free survival of patients. In addition, inhibition of FAK and ERK1/2 reversed CLCA2 silencing-induced tumor cell migration. Furthermore, inhibitors against chloride channels suppressed NPC cellular migration which could have been enhanced by the presence of CLCA2.
CONCLUSIONS: CLCA2 suppress NPC proliferation, migration, invasion and epithelial-mesenchymal transition through inhibiting FAK/ERK signaling.
摘要：
背景：据报道，CLCA2是一种肿瘤抑制因子，在乳腺癌中失调。然而,其在鼻咽癌肿瘤生长和转移中的作用鲜有报道。在这项研究中,我们研究了CLCA2影响NPC的功能和分子机制。
方法：通过实时PCR（RT-PCR）检测人NPC细胞系和组织中CLCA2的表达，Westernblot和IHC。CLCA2在增殖中的生物学作用，在5-8F中评估了NPC细胞系的迁移和侵袭，S18、S26和SUNE-1细胞。细胞活力，通过MTS在体外评估迁移和侵袭，集落形成和transwell分析，分别。CLCA2在NPC的生长和转移中通过NPC异种移植肿瘤生长在体内进行评估，小鼠肺转移模型和贲门淋巴结转移模型。
结果：CLCA2过表达显著降低增殖，NPC细胞的迁移和侵袭。相比之下,CLCA2的敲低引起相反的效果。CLCA2过表达抑制异种移植瘤生长和肺，在体内的the淋巴结（LN）转移。CLCA2通过抑制NPC细胞上皮间质转化(EMT)和激活FAK/ERK1/2信号通路抑制肿瘤转移。143例鼻咽癌标本的免疫组织化学染色显示CLCA2表达是一个独立的,患者总生存期和无远处转移生存期的有利预后因素。此外,抑制FAK和ERK1/2可逆转CLCA2沉默诱导的肿瘤细胞迁移。此外，针对氯通道的抑制剂抑制了NPC细胞的迁移，而CLCA2的存在可能会增强NPC细胞的迁移。
结论：CLCA2抑制NPC增殖，迁移,通过抑制FAK/ERK信号传导进行侵袭和上皮间质转化。
方法：通过实时PCR（RT-PCR）检测人NPC细胞系和组织中CLCA2的表达，Westernblot和IHC。CLCA2在增殖中的生物学作用，在5-8F中评估了NPC细胞系的迁移和侵袭，S18、S26和SUNE-1细胞。细胞活力，通过MTS在体外评估迁移和侵袭，集落形成和transwell分析，分别。CLCA2在NPC的生长和转移中通过NPC异种移植肿瘤生长在体内进行评估，小鼠肺转移模型和贲门淋巴结转移模型。
结果：CLCA2过表达显著降低增殖，NPC细胞的迁移和侵袭。相比之下,CLCA2的敲低引起相反的效果。CLCA2过表达抑制异种移植瘤生长和肺，在体内的the淋巴结（LN）转移。CLCA2通过抑制NPC细胞上皮间质转化(EMT)和激活FAK/ERK1/2信号通路抑制肿瘤转移。143例鼻咽癌标本的免疫组织化学染色显示CLCA2表达是一个独立的,患者总生存期和无远处转移生存期的有利预后因素。此外,抑制FAK和ERK1/2可逆转CLCA2沉默诱导的肿瘤细胞迁移。此外，针对氯通道的抑制剂抑制了NPC细胞的迁移，而CLCA2的存在可能会增强NPC细胞的迁移。
结论：CLCA2抑制NPC增殖，迁移,通过抑制FAK/ERK信号传导进行侵袭和上皮间质转化。
