健脾补肾解毒方通过MALAT1/miR-200s/ZEB1信号轴调控EMT抑制结直肠癌侵袭转移的机制

Colorectal cancer (CRC) is one of the most serious cancers in terms of both incidence and mortality globally. Epithelial-Mesenchymal Transition (EMT) plays important role in the invasion and metastasis of CRC. Previous results demonstrated that lncRNA and miRNA jointly participate in the regulation of tumor EMT, lncRNA-MALAT1 promotes tumor EMT, miR-200c inhibits the EMT of CRC by decreasing ZEB1, whereas its mechanism is unclear. Previous studies revealed that JianPi BuShen JieDu Recipe (JBJR) possess effective effects for the treatment of CRC. Our pilot work showed that MALAT1 and miR-200s have the same targets, and CRC EMT can be suppressed through decreasing MALAT1 after treatment of JBJR. The present project will investigate on the mechanism colorectal cancer EMT promoted by MALAT1 through the regulation of miR-200s/ZEB1 signaling pathway using plasmid, gene silencing and other experimental methods in the three levels of cells, animal and clinical pathology. The effect and mechanism of the invasion and metastasis of CRC suppressing via regulating EMT using MALAT1/miR-200s/ZEB1 signal axis after JBJR treatment, providing important experimental and theoretical basis for revealing the inhibition effect and mechanism of colorectal cancer EMT after treatment of traditional Chinese medicine.

结直肠癌是全球发病率和死亡率最高的疾病之一，上皮-间质转化(EMT)在结直肠癌侵袭转移中起重要作用。研究发现lncRNA和miRNA共同参与调控肿瘤EMT，lncRNA-MALAT1促进肿瘤EMT，miR-200c下调ZEB1抑制结直肠癌EMT。然而，其作用机制尚不清楚。前期临床发现健脾补肾解毒方治疗结直肠癌具有较好疗效。预实验发现MALAT1与miR-200s具有相同靶点；健脾补肾解毒方通过下调MALAT1抑制结直肠癌EMT发生。本课题将从细胞、动物和临床组织病理三个层面，采用质粒转染、基因沉默等实验方法，研究MALAT1通过调控miR-200s/ZEB1信号通路，促进结直肠癌EMT的形成机制；并明确健脾补肾解毒方通过MALAT1/miR-200s/ZEB1信号轴调控EMT抑制结直肠癌侵袭转移的作用及其机理，为揭示中药复方抑制结直肠癌EMT提供重要的实验和理论依据。

结直肠癌是全球发病率和死亡率最高的疾病之一，上皮-间质转化(EMT)是影响结直肠癌侵袭转移的重要因素之一。已有研究表明，多种非编码RNA包括lncRNA和miRNA在内共同参与调控肿瘤EMT。有报道发现，lncRNA-MALAT1促进肿瘤EMT，而miR-200c下调ZEB1抑制结直肠癌EMT。然而，其相互之间的联系和具体作用机制尚不清楚。我们前期临床应用中发现健脾补肾解毒方对治疗结直肠癌具有良好的疗效。生物信息学预测发现，MALAT1与miR-200s具有结合靶点；同时预实验也发现健脾补肾解毒方通过下调MALAT1抑制结直肠癌EMT发生。本课题从细胞、动物和临床组织分析三个层面，采用质粒转染、基因沉默等生物学方法研究发现，MALAT1在结直肠癌患者转移灶组织的表达明显高于原发肿瘤组织，并且MALAT1高表达与高侵袭转移和患者预后密切相关。进一步分子机制研究提示，MALAT1竞争性抑制miR-200s的表达，上调ZEB1，促进结直肠癌发生EMT；另一方面，健脾补肾解毒方通过下调MALAT1，间接上调miR-200s的表达，继而下调ZEB1，最终抑制结直肠癌的EMT和侵袭转移。本研究为揭示中药复方抑制结直肠癌EMT提供了科学实验和理论依据，也为中药复方的进一步开发和临床应用奠定了基础。