| Part I:GLO1 is highly expressed in ESCC tissues and correlated with clinical parameters.ObjectiveEsophageal cancer is one of the most frequently diagnosed invasive malignancies and ranks as the sixth cancer-related mortality worldwide.ESCC is the predominant histologic type of esophageal cancer in China.GLO1 was identified as a potential novel oncogene.In previous study we found that long noncoding RNA(IncRNA)MALAT1 promotes proliferation and metastasis of esophageal squamous cell carcinoma(ESCC),and that the following microarray chip screening of MALAT1 target genes showed that Glyoxalase 1(GLO1)was a potential downstream effector of MALAT1.Overexpression of GLO1 has been shown in various human tumors.However,currently,the role of GLO1 in human ESCC remains unclear.We first determined the expression of GLO1 in ESCC and to explore the clinical significance of GLO1 in ESCC.Methods40 paired ESCC and normal tissues were collected in the Jinling Hospital Affiliated to Southern Medical University.GLO1 mRNA expression was analyzed by qRT-PCR and we analyze the relationship between the clinical parameter and GLO1 expression.ResultsThe expression of GLO1 was notably 2-fold higher than adjacent normal tissues(p=0.0040).GLO1 was over-expressed in tumor tissues in 72.5%(29/40)of all ESCC patients.In order to explore the association between GLO1 expression and clinical pathological characteristics.We divided the 40 patients into two groups(high-expression and low-expression).Association with TNM stage(p=0.024)and lymph nodes metastasis(p=0.031)were analyzed between the two groups.ConclusionGLO1 is highly expressed in ESCC tissues and correlated with clinical parameters.Part II:Mechanism of GLOl promoting malignant ESCCObjectiveGLOl might be an essential effector of lncRNA MALAT1 which promotes ESCC progression.The purpose of this study is to explore the relationship between the expression of GLO1 in esophageal squamous cell carcinoma and the progression of esophageal squamous cell carcinoma,as well as the potential regulation mechanism,it can be identified as a potential therapeutic target for ESCC in the future.Methods1.We designed siRNAs to silence the expression of GLO1 and detect the effect of GLO1 on ESCC cells invasion,proliferation ability,cell cycle and apotosis.2.We conducted nude mice xenograft experiment to investigate the effect of ZIC5 on the growth of NSCLC tumor.3.We analyzed the correlation between the expression levels of MALAT1 in 29 ESCC with high expression of GLO1.After siRNA silence MALAT1 expression in ESCC cell line,qRT-PCR and Western blot method were used to detect the change of GLO1 expression level.Result1.Down-regulation of GLO1 inhibited invasive and malignant capacity in ECA109B and TE13 cells.Knockdown of GLO1 inhibits ESCC cells proliferation and induces G0/G1 cell cycle arrest.2.Mice with sh-GLO1 transfected TE13 cells had smaller tumors compared with those treated with control shRNA since 5 weeks after cell inoculation.The expression intensities of the proliferation marker Ki-67 were weaker in the sh-GLO1 group than in the sh-control group.3.We analyzed the association of expression level between expression of GLO1 and MALAT1 by qRT-PCR in 29 cases with high expression of GLO1 ESCC tissues and found significant correlation between MALAT1 and GLO1(r=0.7217 p<0.01).Furthermore,down-regulation of MALAT1 suppressed the expression of GLO1 by western-blot.ConclusionWe found that Knockdown of GLO1 by siRNA significantly inhibited the proliferation and migration of ESCC cells.In vivo assays showed that knockdown of GLO1 decreased tumor growth.GLO1 was most significantly regulated by MALAT1,which was further verified by tissues and cell experiment,indicating the involvement of MALAT1 in glyoxalase system. |
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