The Studies On Mechanisms Of CPEB1in Regulating Proliferation And Migration Of Bladder Cancer RT112Cells

Background and Objective: Worldwide, bladder carcinoma is common in the malignanttumor. In China, it has the first of urinary reproductive system tumor incidence.Bladder cancer pathogenesis process is a result of polygenic participation, complexfactors and lots of steps of formation.Virus or some chemical carcinogens such asaromatic amine chemical substances on the human body, activate proto-oncogene beoncogene and make tumor suppressor genes inactivation and lead to the occurrence ofcancer.Non-invasive low-level bladder cancer is characterized by lack of chromosome9p/q and activated mutations in FGFR3, PIK3CA and HRAS performance. CPEB1is amember of cytoplasmic polyadenylation element binding proteins, it recruit translationinhibitor to its purpose mRNA3’untranslated region. It is combined with the specificsequence (UUUUUAU) and regulates the polyadenylation of mRNA. As the result, thetranscription of mRNA is regulated by CPEB1. CPEB1in gastric cancer, breast cancerand multiple myeloma cell line is low. The expression decrease of these malignant cellspromotes the invasion and angiogenesis ability. In order to explore the relations betweenCPEB1and bladder cancer cell， proliferation and migration, our experiment throughCPEB1shRNA and Migr1-CPEB1eukaryotic expression vector transfecting RT112cellsand specifically silence and overexpress CPEB1and then observe the affection of CEPB1on RT112cells’ proliferation and migration.Method:1. Firstly we establish shCPEB1interference carrier and CPEB1expressionvector and the transfect RT112cells.2.We use the CFU experiment test CPEB1’ influenceon bladder cancer RT112cell clone formation ability.3.Using AnnexinV staining to detectthe effect of CPEB1on cell apoptosis of RT112cells.4.We use scratch test and transwelltodetect the affection of CPEB1on RT112cells.5.Using Western Blot to test related proteinexpression.Result:1.The CFU results showed the CPEB1overexpressed experimental group has lessnumber of clones than the control group, but the number of clones formed in shCPEB1 interference group is more than in control.2. CCK-8results showed the proliferation ofshCPEB1interference group is faster than the control group while CPEB1overexpressedexperimental group is slower than the control group in proliferation.3. The AnnexinVstaining streaming tests showed both overexpressed and interfered CPEB1group had nodifference in apoptosis rate compared with control group.4. Scratch experiments showedthe migration rate of CPEB1interfered group is slower while CPEB1overexpressedgroup is quicker.5. TransWell results showed CPEB1overexpressed group is quicker thanthe control group in migration, while shCPEB1interfered group compared with controlgroup is reverse.6. WB results showed that CPEB1overexpressed compared with controlgroup, cyclinB1protein expression level is low but p21，c-myc, Hif1-αprotein expressionlevel is high, while the interfered group compared with control group is reverse.Conclusion: CPEB1can inhibit RT112cells to proliferate and to formate clones. It alsocan inhibit RT112cells’ ability to migrate. This may be through regulating p21, cyclinB1,c-myc and Hif1-αgene expression.