The Study Of MiR-10b Targets HOXD10 To Promote Invasion And Metastasis In Esophageal Squamous Cell Carcinoma

Objective By investigating the expression of mi R-10 b and HOXD10 in human ESCC tissues and cell lines and measuring the changes of invasion and migration ability after the esophageal cancer cell lines transfected miR-10 b mimics or inhibitor, the study was to explore the regulation relationship and mechanism between mi R-10 b and HOXD10 in progression and metastasis of ESCC.Methods We investigated the expression of miR-10 b and HOXD10 in human ESCC tissues(30 esophageal cancer tissues and paried normal tissues) and a panel of esophageal cancer cell lines( KYSE30, KYSE70, KYSE140, KYSE150,EC9706,SHEE and SHEEC)with RT-PCR and Western blot,then analyzed the association between mi R-10 b expression level and clinical pathologic features and survival rates. By using bioinformatics software, we speculated that HOXD10 might be one of the target genes of miR-10 b. The esophageal cancer cell lines which expressed low level of miR-10 b were transfected with mi R-10 b mimics and the high level ones were knocked miR-10 b down. Wound healing assays and transwell assays were performed to explore the ability changes of migration and invasion. Using the same methods to inhibited HOXD10,we detected the migration and invasion ability and the expression of tumor invasion-associated protein MMP-9 to explore the regulation mechanism between mi R-10 b and HOXD10 in progression and metastasis of ESCC.Results 1. The expression of mi R-10 b was up-regulated in human ESCC tissues( P=0.017) and its expression level was significantly associated with nodes metastasis(P=0.038). miR-10 b expression level had little associated with clinical pathologic features and survival rates(P>0.05). In esophageal cancer cell lines,the expression of mi R-10 b in EC9706 was the highest than others. Expression of mi R-10 b in SHEE was lower than SHEEC.2. the HOXD10 m RNA expression was down-regulated in human ESCC tissues(P=0.018),the protein expression in ESCC tissues was also lower than paired normal ones. In esophageal cancer cell lines, the HOXD10 mRNA expression in KYSE30,KYSE70,KYSE140 was the higher than the others, and protein expression of HOXD10 in KYSE140 was the highest than others and KYSE150 was the lowest.3. By using bioinformatics software, we speculated that HOXD10 might be one of the target genes of miR-10 b. The expression of HOXD10 protein was up-regulated or down-regulated when miR-10 b was inhibited or overexpressed(P<0.05),but the mRNA level has little changes(P>0.05);4. In wound healing assays and transwell assays,the cell migration and invasion ability was up-regulated or down-regulated when miR-10 b was overexpressed or inhibited. Knocked HOXD10 down,the effect of cell migration and invasion ability was similarity with miR-10 b overexpressed5. MMP-9 was high expression in ESCC tissues.Silencing of HOXD10 in ESCC cells led to the level of MMP-9 up-regulated..Conclusion 1. The expression of miR-10 b was up-regulated in human ESCC and its expression level was significantly associated with nodes metastasis.2. The HOXD10 gene was a direct target of miR-10 b. mi R-10 b promoted esophageal cancer cells invasion and migration by regulating HOXD10.3. mi R-10 b may negatively regulated HOXD10 by up-regulating MMP-9 to promote invasion and metastasis in esophageal cancer.