| Objective: To investigate the effect of lentivirus mediated overexpression of Eca109WISP2 on esophageal carcinoma in nude mice Methods:1.The effect of WISP2 overexpression on the proliferation of human esophageal carcinoma cell line Eca109 was detected by MTT cell proliferation assay;2.The effect of WISP2 overexpression on invasion and metastasis of esophageal carcinoma cells was observed by Transwell chamber invasion assay;3.The effect of over expression of WISP2 on the metastasis of esophageal cancer cells was observed by cell scratch assay;4.To extracted the total RNA gene to detect the expression of WISP2 and E-cadhein gene in each group by real time PCR method5.To establish all the nude mice models of esophageal carcinoma which were divided into three groups,Eca109 control group,Eca109-empty vector group,Eca109-WISP2 expression.To choose the right axillary injection amount of cell suspension,regular measurement of tumor growth in volume,record and process data,and draw the tumor growth curve;21 days later,the nude mice were sacrificed by cervical dislocation,take out the tumor tissues and then.to measure the final weight of each tumor;6.Take out a moderate amount of nude mice tumor tissue,to extracted the total RNA gene in tumor tissue and detect the expression of WISP2 and E-cadhein gene in each tumor tissue by real time PCR method7.Take out a moderate amount of nude mice tumor tissue,to extracted the total protein in tumor tissue and detect the expression of WISP2 and E-cadhein protein in each tumor tissue by Western Blotting method8.Immunohistochemical method was used to detect the expression of WISP2 and E-cadherin protein in the transplanted tumor tissues of each group,and to explore therelationship between them;Results:1.In the MTT cell proliferation assay,WISP2 overexpression stable cell strain(Eca109-WISP2 overexpression group)cell proliferation was significantly lower than that of Eca109-control group and Eca109-vector group,p < 0.05,there were statistically significant differences;but as for cell proliferation there was no significant difference between normal group and negative control group,p>0.05.2.In the Transwell invasion experiment WISP2 overexpression stable cell strain invasion was significantly lower than that of blank group and vector group cells,p<0.05,there were statistically significant differences;but the blank group and empty vector group cell invasion ability compared to the p>0.05,there were no significant difference.3.In the cell scratch experiments,overexpression of WISP2 in metastasis of stably transfected cell lines was significantly lower than control group and vector group cell,p<0.05,there were statistically significant differences;but to compared the control group and empty vector group cell metastasis p>0.05,the difference was not statistically significant.4.In real time PCR experimentsthe of the cell,the WISP2 stable overexpression group WISP2,E-cadherin gene expression level was significantly higher than the blank group and empty load group,and the difference was statistically significant,p<0.05.5.The nude mouse model of esophageal cancer was constructed successfully,the tumor formation rate was 100%,WISP2 overexpression stable cell tumor growth volume was significantly lower than control group and empty vector group,p<0.05,the differences were statistically significant,but to compare the control group with empty vector group,p>0.05,there were no significant difference;The final weight of WISP2 over expressing stable cell group was significantly lower than that of blank group and blank group,the difference was statistically significant,and the difference between the blank group and the blank group was not statistically significant6.In real time PCR experiments of the tumor tissue,WISP2 stably over expressedtumor tissue,WISP2,E-cadherin gene expression levels were significantly higher than those in the blank group and empty body group,and the differences were statistically significant7.The Western Blotting experimental results of tumor tissue in mice showed that the high expression of WISP2 in wisp 2protein stably transfected group sustained,significantly higher than the control group and empty vector group,and the difference is statistically significant,there was no significant difference between the control group and empty vector group.At the same time WISP2 stable expression of E-cadherin was significantly higher than the control group compared and the control vector,the differences were statistically significant,but the difference between the control group and the empty vector had no statistical significance8.The Immunohistochemistry results of the nude mice tumor showed that the expression of WISP2 and E-cadhein protein in the WISP2 stable group was significantly increased than they were in the blank group and the blank group,and the difference was statistically significant Conclusions:1.The stable overexpression of WISP2 can significantly decreased the proliferation of esophageal cancer cells;2.The stable overexpression of WISP2 significantly inhibited invasion and migration ability of esophageal cancer cells;3.The stable overexpression of WISP2 in Eca109 cells in the nude mice tumor,and then which can significantly inhibit tumor growth and body weight of the nude mice4.In vivo experiments confirmed the stable overexpression of WISP2 can promote E-cadherin,the increase of E-cadhein can reverse the EMT,reduce the invasion ability of tumor cell proliferation,suggesting that WISP2 may become a new target for biological therapy in the treatment of esophageal cancer。... |
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