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目的探讨MAP1LC3B在结直肠癌(CRC)中的表达与侵袭转移及自噬的作用机制。方法采用免疫组织化学方法检测92例结直肠癌组织中MAP1LC3B的表达情况,结合临床病理及随访资料进行分析。体外构建慢病毒pLKO.1-MAP1LC3B-siRNA干扰质粒,转染HCT-116细胞株,对照组转染空质粒;采用MTT和侵袭小室实验检测细胞增殖侵袭能力。免疫荧光观察细胞形态。定量RT-PCR和Western blot方法检测MAP1LC3B、mTOR、ATG5和HIF-1α蛋白表达水平。结果 MAP1LC3B高表达存在于82.6%(76/92)癌组织,高于癌旁组织中的阳性率11.95%(11/92),差异有统计学意义(P<0.05);MAP1LC3B蛋白高表达与肝转移、TNM分期和不良预后具有显著相关性(P<0.05);抑制MAP1LC3B表达降低HCT-116细胞侵袭能力;并促进细胞发生自吞噬;下调mTOR、ATG5和HIF-1α蛋白表达水平,差异有统计学意义(P<0.05)。结论 MAP1LC3B是CRC新的预后指标;MAP1LC3B调控mTOR、ATG5和HIF-1α表达介导自吞噬的作用分子机制在CRC侵袭转移中起重要作用。
Objective To investigate the expression of MAP1LC3B in colorectal cancer (CRC) and its mechanism of invasion, metastasis and autophagy. Methods The expression of MAP1LC3B in 92 cases of colorectal cancer tissues was detected by immunohistochemistry. The clinicopathological features and follow-up data were analyzed. The lentiviral plasmid pLKO.1-MAP1LC3B-siRNA was constructed in vitro and transfected into HCT-116 cell line. The control group was transfected with empty plasmid. Cell proliferation and invasion were detected by MTT assay and invasion chamber assay. Immunofluorescence observation of cell morphology. Quantitative RT-PCR and Western blot were used to detect the expression of MAP1LC3B, mTOR, ATG5 and HIF-1α. Results The high expression of MAP1LC3B was found in 82.6% (76/92) of cancerous tissues, which was significantly higher than that in paracancerous tissues (11.95%, 11/92) (P <0.05). MAP1LC3B protein was highly expressed in hepatocellular carcinoma Metastasis, TNM stage and poor prognosis (P <0.05). Inhibition of MAP1LC3B expression reduced the invasion ability of HCT-116 cells and promoted cell autophagy. The expressions of mTOR, ATG5 and HIF-1α were down-regulated Significance (P <0.05). Conclusion MAP1LC3B is a new prognostic indicator of CRC. The molecular mechanism of MAP1LC3B regulating the expression of mTOR, ATG5 and HIF-1α plays an important role in CRC invasion and metastasis.