Lentivirus-mediated shRNA Targeting CNN2 Inhibits Hepatocarcinoma in Vitro and in Vivo.
Lentivirus-mediated shRNA Targeting CNN2 Inhibits Hepatocarcinoma in Vitro and in Vivo.
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慢病毒介导的靶向 CNN2 的 shRNA 在体外和体内抑制肝癌
DOI：
10.7150/ijms.21113
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发表时间：
2018
影响因子：
3.6
通讯作者：
Zhou S
中科院分区：
文献类型：
--
作者：
Kang X;Wang F;Lan X;Li X;Zheng S;Lv Z;Zhuang Y;Zhao Y;Zhou S
目的：肝细胞癌是最常见的恶性肿瘤之一，死亡率高。我们先前的研究表明，在肝细胞癌组织中，尤其是在转移性肝癌组织中，钙蛋白2(CNN2)的表达上调。为了更好地了解CNN2在肝细胞癌中的作用，采用RNA干扰技术(RNAi)探讨其在肿瘤生长和转移中的作用。方法：将慢病毒介导的CNN2-shRNA导入SK-HEP-1细胞，采用实时定量聚合酶链式反应(qRT-PCR)、免疫印迹法(WB)、Transwell法、四甲基偶氮唑蓝比色法和流式细胞术分别检测CNN2基因表达、细胞迁移、侵袭、增殖和细胞周期的变化。将慢病毒CNN2shRNA导入SK-Hep-1细胞裸鼠体内，观察CNN2shRNA对肿瘤生长的影响。采用苏木精-伊红染色、原位末端标记法、免疫组织化学技术分别检测移植瘤组织的组织病理学、细胞凋亡率、总蛋白及其相应的磷酸化蛋白MEK1/2、ERK1/2、AKT的表达。结果：我们的研究表明，CNN2shRNA能有效下调CNN2mRNA和蛋白的表达，抑制细胞增殖，使细胞周期停滞在S期，减少细胞的迁移和侵袭。带有CNN2下调的SK-HeP-1细胞显著抑制了裸鼠体内的肿瘤生长。异种移植瘤组织具有典型的肿瘤特征，shRNA组和对照组均未检测到细胞凋亡。各组裸鼠均未发现转移性肿瘤。随着CNN2蛋白表达下调，除PACK、AKT、MEK1/2和ERK1/2外，pMEK1/2和pERK1/2蛋白表达均明显下调。结论：CNN2可能通过MEK1/2-ERK1/2信号通路在肿瘤生长和转移中发挥重要作用。我们的研究表明，CNN2可能是肝癌分子靶向治疗的潜在靶点。
Objective: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors with a high rate of mortality. Our previous study shows the expression of calponin 2 (CNN2) is up-regulated in hepatocellular carcinoma tissues, especially in metastatic ones. To better understand the role of CNN2 in HCC, RNA interference (RNAi) was used to explore its role in tumor growth and metastasis. Methods: Lentivirus-mediated CNN2-shRNA was transfected into SK-hep-1 cells, and the efficacy of CNN2 expression, cell migration, invasion, proliferation and cell cycles were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR), Western blot (WB), Transwell assay, methyl thiazol tetrazolium assay and flow cytometry, respectively. SK-hep-1 cells transfected with Lentivirus-CNN2 shRNA were xenografted in Balb/C nude mice to explore the effect of CNN2-shRNA in tumor growth. Xenograft tumor tissues were examined for their histopathology, cell apoptosis, the expression of total protein and their corresponding phosphorylated protein of MEK1/2, ERK1/2, AKT, by hematoxylin and eosin stain (H & E staining), TUNEL assay, immunohistochemical technique, respectively. Results: Our research shows it is evident that CNN2 shRNA can effectively down-regulate the expressions of CNN2 mRNA and protein, inhibit cell proliferations, arrest cell cycles at the S phase and reduce cell migration and invasion. SK-hep-1 cells with CNN2 down-regulation have markedly attenuated tumor growth in nude mice. Xenograft tumor tissues have displayed typical tumor characteristics and no apoptosis is detected in shRNA group or in control group. No metastatic tumor was found in any group of nude mice. With CNN2 protein down-regulation, the protein of pMEK1/2 and pERK1/2 are effectively down-regulated, except pAKT, AKT, MEK1/2 and ERK1/2. Conclusions: CNN2 plays an important role in tumor growth and metastasis, possibly through MEK1/2-ERK1/2 signaling pathway. Our study illustrate that CNN2 might be a potential target in HCC molecular target therapy.
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影响因子：
9.7
作者：
Pogribny, Igor P.;Rusyn, Ivan
通讯作者：
Rusyn, Ivan
影响因子：
4.8
作者：
Huang, Qi-Quan;Hossain, M. Moazzem;Jin, Jian-Ping
通讯作者：
Jin, Jian-Ping
影响因子：
--
作者：
Sui, Yanxia;Zheng, Xiaoqiang;Zhao, Dongli
通讯作者：
Zhao, Dongli
影响因子：
4.8
作者：
Tang, J;Hu, G;Sukhatme, VP
通讯作者：
Sukhatme, VP
影响因子：
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作者：
Chen, Yuyan;Liu, Qingqing;Chen, Xudong
通讯作者：
Chen, Xudong