The Effect And Potential Mechanism Of Cp And Sp On Hepatoma Carcinoma Smmc-7721 Cell

Hepatic cell careiroma (HCC) is one of the most frequent malignant tumors in China. It often propagates within liver at the early stage. Because of the lacking of specific symptoms and signs, it is difficult to be diagnosed at early stage. Furthermore, its development is very rapid; many patients miss the chance of surgery treatment. The most of patients are not sensitive to radiotherapy and Systemic chemotherapy. All these above-mentioned facts contribute to high death rate of the patients shortly after the onset of the disease.Currently used chemotherapeutics for cancers primarily are chemically synthesized. Their disadvantages include high medical expense and side effects, such as, damage on normal cells and immunological inhibition. Researchers have been paid a great attention on searching low cost, less side effects, but high effectiveness drugs for cancer treatment. The natural medicinal plants are the most attractive candidates. Some of them could effectively inhibit tumor growth and promote tumor cell apoptosis. Their side effects are much less than the traditional chemotherapeutics.Kudzu (Pueraria lobata) (Willd) can be used as food and drug. FAO expert orecasted kudzu probably become 6^th food supplies crop. There were lots of Pueraria lobata (Willd) Ohwi growing in DA BIE Mountain, The exploit of kudzu in the ascendant. It had kinds of pharmacological action. In domestic research, Du Deji had observed Radix Puerariae's anti-tumor action against many kinds of cancers in vitro. Up to date, little is known about the inhibitory effects of CP and SP on SMMC-7721 cell. The report is limited regarding the correlation between the CP components and their pharmacodynamics.Apoptosis is an active death procedure programmed by genes.The imbalance of cell proliferation, apoptosis and cell cycle leads to tumor.Therefore, this study was designed to explore the effect and its potential mechanism of CP and SP on SMMC-7721 cell through cell culture in vitro at three aspects of anti-proliferation, inducing apoptosis and regulating cell cycle.ObjectivesThe objective of this research were to observe the influence of CP and SP on SMMC-7721 cell proliferation viability, cell cycle distribution and apoptosis rate and to explore its potential mechanism of anti-proliferation.Methods1. SMMC-7721 cell growth inhibiting rate was detected by MTT assay at different concentrations of CP and SP after 24,48, 72hours.2. Apoptosis was observed by TdT-mediated x-dUTP nick end labeling.3. cell cycle distribution were analyzed by flow cytometry.4. The potential molecular mechanisms of anti-proliferation, inducing apoptosis and regulating cell cycle in SMMC-7721 cell were studied at protein expression level of Bax,Bcl-2,CyclinD1 and P27 using immunohistochemistry and flow cytometry.Results1. Both CP and SP effectively inhibited SMMC-7721 cell proliferation. The effects were a dose-dependent and time-dependent. The suppression effect of CP was stronger than that of SP under same condition.2. The quantitative analysis showed the apoptosis rate induced by CP and SP were 17.04% and 5.69% respectively. There was significant difference between CP and control group.3. The effect on cell cycle showed that CP and SP could change the the cell cycle. The percentages of G₀/G₁ phase cell numbers were 75.50% and 60.50% respectively, while those of S phase were 12.95% and 20.65% respectively. There was significant difference between treated and control group.4. The expression ratio of Bax to Bcl-2 was significantly higher in CP and SP groups than that in the control, of which, the ratio value in CP group was significantly higher than the in SP group.5. The effects of CP and SP on SMMC-7721 showed that the P17 and Cyclin D1 proteins were significantly enhanced and declined respectively, of which, the effect of CP was stronger than that of SP.Conclusions1. Both CP and SP inhibited SMMC-7721 tumor cell proliferation. The inhibiting effect of CP was stronger than that of SP.2. SMMC-7721 tumor cell treated with CP and SP were remarkably arrested at G₀/G₁ phase.3. CP induced SMMC-7721 tumor cell apoptosis through up-regulation of expression proportion of Bax/Bcl-2, and changed in the express in mRNA of Bax and Bcl-2.4. Both CP and SP changed SMMC-7721 tumor cell cycle through down-regulation of CyclinDi and up-regulation of P27 protein. Thus, SMMC-7721 tumor cell were accumulated in G₀/G₁ phase.