The Effects Of Huaier Extract On The Proliferation And Expression Of P-YAP In Hepatocarcinoma HepG-2Cells

Hepatocarcinoma (HHC)is a world-wide malignant tumor.HHC is nonsensitive tochemotherapy,clinical reports show that the partial response to chemotherapy of HHCis less than20%up to now. The efficacy of targeted antineoplastic drug on HHC, suchas sorafenib doesn’t improved significantly therapeutic efficacy in HHC. So theefficient drug against HHC is anxiously expected all the time. Huaier extract is atraditional Chinese antineoplastic medicine extracted from Trametes robiniophilaMurr.Clinical data shows it can not only inhibit tumor growth but also improve theliving quality of the patients with tumor. Basic studies illustrate multi-target effects ofHuaier extract on tumor, such as DNA and RNA synthesis inhibition, cell proliferationinhibition, antiangiogenesis, and apoptosis induction. Hippo-YAP pathway plays animportant role in cell growth, apoptosis and organ size. YAP is a downstream effectorin the Hippo-YAP pathway, its abnormal expression is closely related to tumor.TheP-YAP protein can inhibit cell proliferation, promote apoptosis.Objective:This study is designed to investigate the effects of Huaier extract on theproliferation and expression of P-YAP in HepG-2cells and discusses the inhibitingmechanism of Huaier extract in HHC.Methods:1. Inverted microscope was used to observe cell morphology of HepG-2cells after24h and48h impactence of Huaier extract on the cells with differentconcentrations respectively.The experiment was repeated three times.2. CCK-8method was used to analyse proliferation of HepG-2cells after12h,24h,and48h impactence of Huaier extract on the cells with different concentrationsrespectively.The experiment was repeated three times.3. FCM technique was used to analyse apoptosis of HepG-2cells after24h and48himpactence of Huaier extract on the cells with different concentrations respectively.Theexperiment was repeated three times. 4. Western Blot was used to analyse the protein expression of P-YAP in HepG-2cells after11h,12h,24h impactence of Huaier extract on the cells with differentconcentrations respectively.The experiment was repeated three times.Results:1. Inverted microscope analysis shows that Huaier extract inhibitssignificantly HepG-2cells growth in dose and time dependent ways.2. The CCK-8analysis shows that Huaier extract inhibits significantly HepG-2cells proliferation in dose and time dependent ways.3. The FCM analysis shows that Huaier extract promotes significantly HepG-2cells apoptosis in dose and time dependent ways.4. Western Blot analysis shows that Huaier extract increase P-YAP expression ofHepG-2cells after1h impactence on the cells in dose dependent ways,However, fail toeffect the expression of P-YAP after12h impactence,and yet decreases significantly theprotein expression of P-YAP after24h impactence on HepG-2cells in dose dependentways.Conclusion: Huaier extract inhibits significantly HepG-2cells proliferation,promotes the cells apoptosis in dose dependent ways.Increasing protein expressionof P-YAP may be the early phase inhibiting mechanisms of Huaier extract in HHC,With time conditions change, Huaier extract may activate different anti-tumor mechanisms.Huaier extract is a promising medicine on HHC treatment.