Effect And Mechanism Of Connexin43Overexpression On Growth Migration And Apoptosis Of U251Cell

Gap junctions are specialized cell–cell junctions that directly link the cytoplasmof neighboring cells. They mediate the direct transfer of metabolites and ions fromone cell to another. Expression of astrocytic gap junction protein, connexin43(Cx43),is often reduced in astrocytomas, the most common neoplasia of the central nervoussystem (CNS) in adults. Supported by a number of studies, the global decrease ofCx43expression appears to be advantageous for the growth of glioma cells. Our studyis based on over-expressing Cx43to glioma U251, researching the effect andinfluence of grow, migration and apoptosis when over-expressing Cx43to gliomaU251by cell migration assay, RT-PCR and Western Blot,etc. We will provide newthinking and method for treating glioma.1. Affect of Cx43for cell proliferationOur study use normal U251and Cx43-U251cell,counted and planted them in24-well plates for10%FBS IMDM medium,respecitivily. The cells are cultured for7days in all and taken average index number for everyday time when separate the cellsfrom the plate.Experimental result find that U251and Cx43-U251cells grows fasteron4-5days, steady on6-7days.Two types cells have no distinction for grow on1-4days, but U251cells have significant difference for grow compare withCx43-U251cells on5-7days.Test results show that over-expressing Cx43to U251cell may influence the growof cells.The affect of Cx43prevent cell grow could be associated with cell cycleregulatory proteins. Regrettably，we are not clearly understand that which manner forCx43could interact with cell cycle reulatory proteins. Cx43may prevent glioma cellsgrow which give us a new therapy for treating with gliomas.2. Affect of Cx43for cell migrationFor previous part, we have tested the affect of Cx43for cell grow.Cx43seemingly prevent glioma grow. On this part, we want to test the affect of Cx43forcell migration.It finds that U251and Cx43-U251cells in Wound Healing assay,Cx43-U251cells have more directionality for repairing the scratch and normalU251cells have significant difference for repair capacity and movement performancecompared with Cx43-U251cells. Except two different points above, over-expresionof Cx43can make cell skeletons changed during the cells migration for repairingcapacity. Different with the normal U251cells, Cx43-U251cells’ skeletons showextend. Infiltrative growth of gliomas is hard difficulty for clinical therapy, Cx43mayinteract with cell skeletons proteins to affect cells migration. Our results indicate thatover-expression of Cx43could repair capacity faster and enhance capacity of cellmigration.3. Affect of Cx43for cells apoptosisAt present GBM patients generally receive a multimodal approach of surgicalresection, radiotherapy and chemotherapy. TMZ is currently the most effectivechemotherapeutic demonstrating excellent CNS bioavailability. We use1000uM TMZto induce cell apoptosis in experiments. The test establish2groups, normal U251cellgroup and Cx43-U251cells groups. Testing the level of Cx43, Caspase-3andBax/Bcl-2expression. All the experimental results find that Cx43-U251cells groupshave a lower ratio of Bax/Bcl-2compare with normal U251cells groups（p<0.05）.Results indicate that Cx43may affect cell apoptosis by mitochondria pathway.In conclusion, over-expression of Cx43can make a series of effect for U251cells.Cx43have role to prevent U251cell proliferation, promote cells migration andresist apoptosis by TMZ induced.