The Role Of MiR-630 In Colorectal Cancer,and Its Related Mechanism Research

The first part: Expression of miR-630 in colorectal cancerObjective: Different expression of miR-630 in cancer tissue and normal mucosa were detected to analyze the possible relationship between pathogenesis and pathology.Methods: Forty five pairs of tumor and normal mucosa from colorectal cancer patients were collected.Total RNA of the two kinds groups were extracted respectively.As consequence,cDNA were made by reverse transcription.The relative expressions of miR-630 were measured by real-time fluorescent quantitative PCR technology.The results were analyzed by software of SPSS17.0.The statistically difference was considered to be existed by using paired student test and chi-square test when P < 0.05(double side).Result: 1.It makes significant statistic difference in expression between the two groups in the forty five pairs of samples(P<0.001).Expression of miR-630 in tumor tissue was higher than normal.2.Expression of miR-630 in different TNM stage varies;Expression in T3,T4 were higher than T1,T2(P=0.012),tissues with lymphatic metastasis were higher than those not ones(P=0.002),advanced stagesexceed early stage(P=0.027).Conclusion: It shows that expression of miR-630 in tumor tissue were higher than normal mucosa,implying the participation of miR-630 in tumorigenesis and development of colorectal cancer.The second part: The role of mi R-630 in cell lines SW480 and mechanism researchObjective: 1.To clarify the probable tumorigenesis and tumor development functions of mi R-630 in colorectal cancer,proliferation,migration,aggression in Cell strain SW480 were investigated.2.The regulative relationships between MiR-630 and CDKN2 B gene were explored to discuss the possible mechanism.3.Experiments of tumor in vivo were conducted to explore the effects of mi R-630.Methods: 1.Stable cell strains transfected with anti-miR-630(mi R-630 inhibitor)were screened to compare with the negative control on expression.2.Investigations on the cell behavior were demonstrated by experiment to find the different characteristic of proliferation,cell cloning,invasion and metastasis.3.MiR-630’s potential target genes were picked out by TARGETSCAN.The relationship between miR-630 and CDKN2 B was confirmed by western blot which verified the expression of CDKN2 B in Anti-miR-630(miR-630 inhibitor and the negative control.The same wasdone in tumor tissue.4.Stable transfected cell strains were implanted into nude mice subcutaneously to observe the tumorigenesis influences of miR-630 in vivo.The results were analyzed by software of SPSS17.0.The statistically difference was considered to be existed by using Two sample T test when P < 0.05(double side).Result: 1.The expression of mi R-630 in the Anti-mi R-630 declined obviously,lower than negative control,which have a statistically significant differences(P<0.05).2.The cell proliferation,invasion and metastasis in Anti-miR-630 were inferior to the negative control.3.The expression of CDKN2 B improved in Anti-mi R-630 cell strains than negative control,while the opposite was happened in the tumor tissues.4.The weight and size in Anti-miR-630 were below to the negative control.Conclusion: In a word,mi R-630 may be associated with the cell proliferation,invasion and metastasis of colorectal carcinoma concerning the result of implanted tumor subcutaneously in nude mice and the fact that Anti-miR-630 can impress the ability of the cell proliferation,invasion and metastasis;A mechanism to promote the development of colorectal cancer occurrence perhaps was obtained by downregulating the expression of CDKN2B。...