The Clinical And Experimental Study Of Jianpi Huayu Therapy On Hepatocellular Carcinoma

ObjectiveIn China hepatocellular carcinoma (HCC) is characterized by high incidence and poor prognosis. High invasiveness and potential to metastasis is one of the most important characteristics of HCC. Unfortunately, long-term survival after treatment for HCC is still unsatisfactory. Most of HCC patients failed to receive potentially curative treatment, such as hepatic resection for HCC because of tumor metastasis. Molecular targeted drugs (Sorafenib) and chemotherapy were used as systemic therapy for metastatic HCC in modern western medicine. Low Objective response rate, expensive price, and significant side effects resulted in unsatisfactory treatment effect. Traditional Chinese Medicine possessed some of the features and advantages for advanced HCC. The theory of "Jianpi Huayu Therapy" is rooted in "Synopsis of Golden Chamber""Jianpi Huayu Therapy" is one of the most important therapies on HCC. However, most of the previous studies on the anticancer mechanisms of "Jianpi Huayu Therapy" aimed at inhibition of proliferation or induction of apoptosis. Epithelial mesenchymal transition (EMT) is a process by which epithelial cells lose their epithelial charateristics, cell polarity and cell-cell adhesion, and acquire invasive properties and stem cell-like features. Recent studies found EMT provides the basis for invasion and metastasis of HCC. Smad3was the control point of TGF-β/Smads pathway, which was the most important pathway involved in EMT. Therefore, investigating the clinical effect of "Jianpi Huayu Therapy" in the treatment of hepatocellular carcinoma (HCC), and investigating the impacts of Jianpi Huayu therapy on EMT of HCC cell will reveal the anticancer mechanism of "Jianpi Huayu Therapy" on HCC, and lead to a novel therapeutic strategy and theory basis for Chinese medicine treatment on HCC. Such scientific problems was worthy of further exploration and was of great practical significance. The aim of this study is to investigate the clinical effect of hepatectomy combining with "Jianpi Huayu Therapy" in the treatment of HCC, and to investigate the impacts of Jianpi Huayu therapy on EMT of HCC in nude mice via Smad3protein regulation.Methods1)Clinical study:One hundred and twenty HCC patients were randomized into hepatectomy combined with "Jianpi Huayu Therapy" group (treatment group, n=60) and hepatectomy alone group (control group, n=60). Disease-free survival (DFS) and overall survival (OS) were used as primary end-point to evaluate the treatment effect. The liver function at one week after operation, complication, the average days of hospitalization and performance status (PS) at one month after operation were evaluated.2)Experimental study:Forty nude mice with HCC cell line MHCC97-H orthotopic xenografts were randomized to two groups:"Jianpi Huayu Therapy" Chinese medicine intragastric administration group (treatment group, n=20) and sterile saline intragastric administration group (control group, n=20). The numbers and size of tumor lesions in the liver and lung were observed respectively. The protein level of Smad3, E-cadherin, N-cadherin and vimentin in40liver cancer samples were determined by immunohistochemistry method respectively.Results1)Clinical study:There was no significant difference in baseline data between two groups (P>0.05). No peri-operative mortality occurred in two group. Fourteen cases (23.3%) developed post-operative complications in treatment group, twelve cases (20.0%) in control group. There was no significant difference between two groups (x2=0.196,P=0.658). Alanine aminotransferase (ALT) at one week after operation in treatment group was lower than in control group (t=2.053, P=0.042). There were no significant differences in the other indexes of liver function at one week after operation between two group. The average days of hospitalization in treatment group decreased by0.9day than in control group (t=2.146, P=0.034). No significant differences occurred in the average scores of PS at one month after operation between treatment group (1.05±0.47) and control group (1.17±0.46),(t=1.183, P=0.169). At the time of censor,104patients (86.6%) developed recurrence. The1-,3-, and5-year disease-free survival rates and median disease-free survival for all the patients were77.4%,26.3%,9.0%and25.6months (range,6.0-68.0), respectively.48patients in the treatment group and56patient in the control group, respectively,(x2=4.615, P=0.032). The median DFS and1-3-and5-year DFS rates for treatment group and control group were28.7months,78.2%,29.2%,14.3%and22.6months,75.0%,23.3%,6.4%, respectively (x2=4.017, P=0.045).101patients had died at the time of censor. The1-,3-, and5-year overall survival rates and median survival for all the patients were97.5%,76.4%,40.5%and51.2months (range,10.0～72.0), respectively. The median OS,1-,3-and5-year OS rates for treatment group and control group were52.6months,98.3%,78.0%,43.6%and49.8months,96.7%,74.7%,37.4%, respectively (x2=3.921,P=0.048).2)Experimental study:Forty nude mice with HCC cell line MHCC97-H orthotopic xenografts developed HCC lesions in two groups. The size of intrahepatic tumor [(2180±690)mm3] of treatment group was smaller than that of control group [(3151±580)mm3],(t=10.12, P<0.01). The rate of intrahepatic and lung metastasis in treatment group decreased significantly than that of control group, respectively (65%vs.95%, P=0.044;55%vs.90%, P=0.013). The positive rates of Smad3(25%vs.60%, x2=5.013, P=0.025) and N-cadherin (35%vs.70%, x2=4.912,P=0.027) in treatment group were lower than in control group. Positive rate of E-cadherin in treatment group (80%) was higher than in control group (45%),(x2=5.227, P=0.022). Positive rate of vimentin in treatment group was40%, while70%in control group. The difference was not significant (x2=3.636, P=0.057)Conclusion1) The hepatectomy combined with Jianpi Huayu therapy were effect in the treatment of HCC, reduced post-operative recurrence and metastasis and improved the disease-free survival and overall survival of HCC patients.2) Jianpi Huayu therapy inhibited the invasion and metastasis of HCC, potential mechanism was regulating Smad3protein level and inhibiting the EMT of HCC.