Study On The Expression Of MMP-2,MMP-9,TIMP-1,TIMP-2mRNA In Esophageal Squamous Cell Cancer And Their Interaction With HPV16E6,HPV16E7,HPV16E6/E7

Objective:to explore the expressional difference of MMP-2,MMP-9,TIMP-1, TIMP-2mRNA between esophageal squamous cell cancer(ESCC) tissues and matched para-carcinoma normal esophageal mucosa,studying the correlation among these four genes in ESCC simultaneously; finding out whether any difference exists or not between different nationalities on the expression of MMP-2, MMP-9, TIMP-1, TIMP-2mRNA in ESCC tissues, whether a regulatory relationship could be found between HPV16E6,E7,E6/E7and expression of MMP-2,MMP-9,TIMP-l,TIMP-2mRNA, furthermore, to explore whether histone deacetylase(HDAC) involved in the expression of MMP-2, MMP-9, TIMP-1, TIMP-2mRNA. Methods:100removed fresh esophageal cancer tissues and matched para-cancer normal tissues were selected to be used as experiment spicemen, detect the expression of MMP-2,MMP-9,TIMP-1,TIMP-2mRNA in both cancer tissues and matched normal esophageal mucosa through reverse transcription polymerase chain reation(RT-PCR),detect randomly5cases from100spicemen protein expression of those four genes by Western-blot. Construction gene carriers of HPV16E6,HPV16E7and HPV16E6/E7,transfection these three carriers into the cell line KYSE450,which is a esophageal squamous cell carcinoma cell with HPV negative, then detect the expression of those four genes in the transfected cell KYSE450. detect the expression of those four genes in KYSE450after adding TSA, which is a kind of HDACi,to the culture of KYSE450. Results:the mRNA expressions of MMP-2,9and TIMP-1,2in cancer tissues are higher than them in matched normal esophageal mucosa, those expressions in Hans are higher than them in Kazakhs, those expression in invasive depth of T3+T4are higher than them in invasive depth of T1+T2, there is no different expressional rate of those four genes between lymph node metastasis positive group and negative group, there is no different expression rate of those four genes between different genders, ages, differentiations of tumor cells either. A positive correlation exists between MMP-2mRNA and MMP-9mRNA, no correlation exists among MMP-2mRNA, TIMP-1mRNA and TIMP-2mRNA, a positive correlation exists between MMP-9mRNA and TIMP-1mRNA, TIMP-2mRNA respectively. The expression of MMP-2mRNA is very low in each transfected KYSE450cells, the expression of MMP-9mRNA is higher in both control group and pEGFP-N1transfected group than it in pEGFP-N1-E6, pEGFP-N1-E7, pEGFP-N1-E6E7transfected groups. There is no different expression of TIMP-1mRNA between pEGFP-N1-E6and control groups, a different expression of TIMP-1mRNA exists between pEGFP-N1-E7and control groups, a different expression of TIMP-1mRNA exists between pEGFP-N1-E6/E7and control groups too. A different expression of TIMP-2mRNA exists between pEGFP-N1-E6and control groups, there is no different expression of TIMP-2mRNA between pEGFP-N1-E7and control groups, a different expression of TIMP-2mRNA exists between pEGFP-N1-E6/E7and control groups too. There is no different expression of MMP-2mRNA, MMP-9mRNA, TIMP-1mRNA between TSA induced group and control group, the expression of TIMP-2mRNA is lower in TSA induced group than in control group. Conclusions:MMP-2,9and TIMP-1,2maybe involved in the carcinogenesis and develop-ment of ESCC, MMP-2,9and TIMP-1,2maybe be related to the invasive growth of ESCC, but maybe not relate to genders, ages and differentiation of tumor cells. A different expression of those four genes maybe exist between Hans and Kazakhs. A positive correlation exists between MMP-2mRNA and MMP-9mRNA, no correlation exists among MMP-2mRNA, TIMP-1mRNA and TIMP-2mRNA, a positive correlation exists between MMP-9mRNAand TMP-1mRNA, TIMP-2mRNA respectively. The carriers of HPV16E6, HPV16E7and HPV16E6/E7were constructed successfully, the transfection of KYSE450was successfully too. The regulatory relationship maybe not exist between HPV16E6, HPV16E7and MMP-2, HPV16E6and HPV16E7maybe suppress the expression of MMP-9, the co-effection of HPV16E6/E7maybe promote the expression of TIMP-1and TIMP-2. The regulatory relationship maybe exist between HDAC and TIMP-2, but the regulatory relationship maybe not exist between HDAC and MMP-2, MMP-9, TIMP-1.