| Objective: Hepatocellular carcinoma, named"King of cancer", is a kind of common malignancy in clinical. Currently, the treatment is based on surgery and chemotherapy, and the latter with adverse reactions such as liver and kidney toxicity is not much satisfactory in the long run. To improve the life quality and survival of patients, the drug with effective and low toxicity is needed. In recent years, the anti-cancer effects of chinese medicine is being recognized gradually. Elemene, developed by China, is a second-line anticancer drug with non-cytotoxic and broad-spectrum, which has been widely used in the treatment of various tumors and the mechanisms are also constantly being studied. To provide more basis for the clinical treatment about chinese medicine, the study observed the antitumor effects of Elemene or combined with 5-FU on H22 tumor cell in mice and explored the possible mechanism.Methods: 35 Balb/C mice (18~22g, 6~8w) were randomly assigned into 5 groups, and made mark on each ear. The prepared cell suspension (4×10~7个/ml) was armpit injected with 0.1ml each mouse. 24 hours later of tumor-bearing, all animals received treatment regimens by intratumoral injection once daily for 10 days: Elemene-higher group (100mg/kg), Elemene-lower group (50mg/kg), 5-Fu group (20mg/kg), 5-Fu (20mg/kg) combined Elemene group (50mg/kg), control group: physiological saline 0.2ml each mouse. The body weights and survival condition of all anmals were recorded through the whole course as the index of drug toxicity. Tumor volume was measured, then calculated the inhibition rate. On day 11, following cancer cell xenograft, all animals were killed by cervical dislocation. Under sterile conditions, tumors were stripped, then photographed. Fixed these specimens for paraffin embedment. And the differences of tumor was observed by HE staining histologically. expression of the pro-apoptotic Bcl-2 family member N by elemene- higher group was detected by immunohistochemical analysis. Results:1. After 3 days of treatment, volumes of tumor of 5-FU group and combined group were less than of control group significantly. To all the 4 groups, there was significant difference with the control group after 5 days(P<0.05). From the photo, all the tumors were observed. Calculated the inhibition rate as average tumor volume of the control group and treatment group differences. It was 10.2%~26.7% in elemene groups and 52.2% in 5-FU group, on the contrary, it could reached 61.0% in combined treatment group.2. Weight loss of the combined and 5-FU groups were more significant. Although there was weight gain due to the increasing tumor volume, compared with control group, the weight of mice in elemene goups were also decreased. Compared with control group, rate of weight loss in elemene group was 0.6%~6.2%, while 5-FU group and combined group was 12.6% and 13.0% and compared with before treatment, rate of weight loss in 5-FU group and combined group was 13.1% and 21.0%.3. In view of diet, activity and hair color, the life quality of the mice in elemene groups were better than others.4. Under the light microscope, the tissue sections by HE staining were observed. Different damage of tumor tissues in each treatment group was seen. In the control group, the cell nuclear was deeply stained, arranged in compact, while in the elemene-higher groups, the nuclear was lightly stained and the cell was swelling, condensation and fragmentation.5. Positive expression of the pro-apoptotic Bcl-2 family member N was brown particles, which was located in cytoplasm. Randomly selected 10 high power fields under the microscope and counted 500 cells. The positive-expression cell in elemene- higher group was 17.2%, which was significant difference from the control group (P<0.05).Conclusion:1. Both elemene and 5-FU can effectively inhibit the growth of H22 tumor.2. Elemene can be used as an adjuvant agent with 5-FU to therapy liver cancer.3. Elemene could inhibit liver cancer by inducing expression of the pro-apoptotic Bcl-2 family member N. |
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