The Effect Of β-catenin On The Proliferation, Invasion, Migration And Apoptosis In Melanoma Cell Of A375

Background：Malignant melanoma is a relatively common malignancy, occur in the skin,it is ahigh degree of malignancy, with an average survival of about30.3months, and theincidence is increasing year by year. Currently, the primary means of treatingmalignant melanoma are surgery and anti-tumor therapy (chemotherapy, radiationtherapy, biological therapy), but their effects are worse. In recent years,Wnt/β-catenin signaling pathway and the development of many cancers are closelylinked was found, but its mechanism is not fully cognitive. The β-catenin is theWnt/β-catenin signaling pathway core protein, and its abnormal accumulation in thecytoplasm is the Wnt/β-catenin signaling pathway activated signs.In this study, wewill silence the expression of β-catenin to observe the change of proliferation,invasion, migration and apoptosis of malignant melanoma A375cells.Objective：To investigate via silencing the expression of β-catenin, the key protein ofWnt/β-catenin signaling pathway, to observe the effect of β-catenin on theproliferation, invasive, migration and apoptosis of A375cells. We aim to acquire newtreatment programs of melanoma.Methods：1. We first make β-catenin-RNAi Lentivirus and β-catenin-negative Lentivirus,later we let two kinds of Lentivirus to infect into A375cells. The cells were infectedwere named A375-RNAi and A375-negative. Then Western blot assay used to detectthe protein expression of β-catenin in all those cells. The cells uninfected wereregarded as the control group.2. The MTT method was to testify the proliferation of A375-RNAi andA375-negative cells.The cells uninfected were regarded as the control group.3. The plate colony assay was used to testify the colony-formation ability of A375-RNAi and A375-negative cells.The cells uninfected were regarded as thecontrol group.4. The Transwell assay was used to discover the capacity of migration andinvasion of A375-RNAi and A375-negative cells. The cells uninfected were regardedas the control group.5. The Sub-G1method of flow cytometry was adopted to investigate theapoptosis rate of A375-RNAi and A375-negative cells. The cells uninfected wereregarded as the control group.Results：1. As showed in Western blot, protein expression of β-catenin in A375-RNAicells is less than that in the other two kind cells. The protein expression inA375-RNAi cells than uninfected A375cells decreased about55.1%(P＜0.05),confirmed that β-catenin-RNAi-LV could effective silence the protein expression ofβ-catenin in A375cells. The expression of β-catenin in A375-negative cells and A375cells have no obvious difference (P＞0.05).2. MTT showed that compared with A375cells the proliferative activity ofA375-RNAi cells was reduced (P＜0.05). The proliferation activity of A375-negativecells and A375cells have no obvious difference (P＞0.05).3. As demonstrated in colony formation assay, the colony-formation ability ofA375-RNAi cells are weaker than A375cells(P＜0.05). But the colony number inA375-negative cells and that in A375cells have no obvious difference (P＞0.05).4. Transwell assay proved that the migration and invasion of A375-RNAi cellswas inhibited after down-regulate the β-catenin expression(P＜0.05). The migrationand invasion of A375-negative cells and A375cells have no obvious difference (P＞0.05).5. Flow cytometry detected that the apoptosis rate of A375-RNAi cells wasobvious increased than A375cells(P＜0.05). But there were no difference betweenA375-negative cells and A375cells (P＞0.05).Conclusion：1. Infected with β-catenin-RNAi-LV could successful down regulate the expression of β-catenin inA375cells.2. Silencing β-catenin expression could lead A375cells proliferationdown-regulated, can inhibit the ability of migration and invasion of A375cells, canweaken the colony-formation ability of A375, can also increase the apoptosis rate ofA375. β-catenin is expected to be a therapeutic target of malignant melanoma.