A Pro-inflammatory Mediator Leukotriene D4 Involved in the Progression of Chronic Hepatitis B and Hepatocellular Carcinoma
|School||Huazhong University of Science and Technology|
|Keywords||chronic hepatitis B(CHB) hepatocellular carcinoma(HCC) hepatitis B virus(HBV) leukotriene D4(LTD4) human hepatocellular carcinoma (HCC) cell line SMMC-7721 Montelukast (MK571)|
Objective: To detect the expression of serum leukotriene D4 in paitents with chronic hepatitis B(CHB) and hepatocellular carcinoma(HCC) in order to investigate the relationship between LTD4 levels and the development of CHB and HCC,and to probe its roles on the progression of HCC.Methods: This study included 21 CHB paitents, 71 HCC patients and 20 controls.Therein, the average age of CHB paitents was 37.6±12.7 years with 16 males and 5 females including light~heavy chronic hepatitis 7 cases and severe hepatitis 14 cases; 71 HCC patients with an average age of 48.9±11.1 years included male 64 cases and female 7 cases, for 62 cases among them accompanying with HBV. In addition, LTD4 levels was detected by Enzyme-linked immunosorbent assay (ELISA), the levels of serum ALT, AST, gamma-GT were examined by Automatic biochemical analyzer, and serum AFP and HBVDNA were measured using Chemiluminescence instrument and Gene amplification instrument,respectively.Results: The serum LTD4 levels of paitients with CHB and HCC were obvously higher than controls (P < 0.01),and its levels gradually declined followed by patients with CHB, HCC with HBV, HCC alone and control group. In CHB group, there was no significant difference between light~heavy hepatitis and severe hepatitis (P > 0.05). Moreover, LTD4 levels were not associated with levels of ALT, AST, gamma-GT and HBV-DNA content. In HCC group, there was significant difference (P < 0.05) between HCC mergered with HBVand HCC alone. Moveover, in metastasis and nonmetastasis group as well as treatment and non-treatment group, there were no statistically difference (P > 0.05). Additonally, there was no correlation between LTD4 and AST,γ-GT or AFP(P > 0.05), but a positive correlation between LTD4 and ALT (P < 0.05). Conclusion: LTD4, as an important proinflammatory mediator, is not only involved in the pathogensis of CHB and could play a role in promoting the progression process from CHB to HCC, but also have some directly effects on HCC cells. Objective: Chronic inflammation plays an important role in carcinogenesis. A pro-inflammatory mediator leukotriene D4(LTD4) not only has been implicated in the pathophysiology of asthma and inflammations, but also in several cancers. However, little is known about the effects of the LTD4 on human hepatocellular carcinoma (HCC). The aim of this study is to investigate the roles of LTD4 on the SMMC-7721 cell proliferation, cell cycle, apoptosis and migration.Methods: By culture human hepatocellular carcinoma cell line SMMC-7721, treated with LTD4 at different concentration and different time and cysteinyl leukotrienes receptor 1 (CysLT1 R) antagonist MK571, cell proliferation were examined by CCK-8 assay in vitro. Then, effects of LTD4 on cell cycle and apoptosis were assessed by flow cytometry. In addition, 8μm Transwell plate was used to detect LTD4 role on the cell migration.Results: The proliferation rate of SMMC-7721 cell was obviously enhanced by LTD4 (P < 0.05), reaching a maximum about 59 percent at a concentration of 1 nmol/L for 48 hour. Furthermore, antagonist MK571 with 10-8-10-6mol/L concentrations prevented partially the roles of LTD4 on cell proliferation by competitive inhibition, while over 10-5mol/L concentration it could significantly inhibit cell growth (P < 0.05). Additonally, SMMC-7721 cell treated by LTD4 within the effective concentrations, the proportion of M1/M2 phase cell declined significantly (P < 0.05) with a dose-dependent manner, promoting cell cycle from M1 from M2; Moreover, LTD4 could apparently inhibit cell apoptosis (P < 0.05) in a dose-dependent, mainly in early-stage apoptosis.Besides, LTD4 also induced cancer cell migration (P < 0.05) and the promotion roles also emerged in the dose-dependent.Conclusion: As an important proinflammatory mediator, LTD4 play roles in the pathogenesis and progression of HCC, indeed. Hence, LTD4 would be as a new potential index to assist the diagnosis of HCC and judgement the prognosis, and its antagonists may also become to new potential drugs chemopreventing the development of chronic hepatitis to HCC in the future.