Study On Interferon-λ Induced Anti-proliferative Activity In Breast Cancer, Esophageal Cancer And Malignant Mesothelioma Cell Lines, And Combinatory Effect With Chem Other Apeutic Agent

Objectives:Currently, worldwide incidence rate of cancer is increasing year by year, the number of people died of cancer is about150millions each year in China. Traditional therapies such as surgery, radiotherapy, chemotherapy sometimes can achieve a clinical cure, but there are still some limitations. In recent years, biotherapy has been clinically recognized because of its treatment of systemic and individualized. biotherapy includes cytokine therapy, treatment of immunocompetent cells, induction of differentiation therapy, gene therapy. Cytokines, including interferon-a, being a new treatment of cancer has been used in clinical and achieved good efficacy. In recent years, the new family members--interferon-λ, was identified, it has similar functions with interferon-a, such as anti-viral, anti-tumor and immune regulatory action. Compared with interferon-a, interferon-λ, has no apparent toxicity and side effects. In this investigation we try to examine the expression of interferon-λ, receptor in breast carcinoma cell lines, esophageal carcinoma cell lines and malignant mesothelioma cell lines, to investigate in which cell lines exists significant proliferation inhibition of interferon-λ, and then to explore the anti-tumor mechanism of interferon-λ. We also use chemotherapeutic agents combining with interferon-λ to see if there has a synergistic inhibition effect. Interferon-λ is a potential therapeutic agent for carcinoma without damaging surrounding tissues.Methods: Firstly, Use PCR to examine the expression of IFN-λ receptor complexes in breast carcinoma231,453MCF-7cells, esophageal carcinoma TE1, TE2, TE10, YES6, T.Tn cells, malignant mesothelioma H2452, H2052H226,211H, H28cells, normal esophagus Het-IA cells. Use MTT to investigate the growth suppressive activity of IFN-λ with these cell lines. Select the cells which express IFN-λ receptor and induce growth suppression after treated with IFN-λ, observe if IFN-λ, produce combinatory anti-tumor effects with chemotherapeutic agents, cisplatin (CDDP) and5-fluorouracil (5-FU). In the end, the respective cleaved forms of caspase3、PARP、caspase8will be analyzed with Western blotting.Results:1IFN-λ, receptors express or not in breast carcinoma cells, esophageal carcinoma cells, normal esophagus cells and malignant mesothelioma cellsWe examined the expression of IL-28Ra and IL-10Rβ genes in fourteen kinds of human carcinoma or normal cell. Esophageal carcinoma TE1, TE2, TE10, YES6, T.Tn cells、normal esophagus Het-IA cells、breast carcinoma453cells and malignant mesothelioma H2052cells expressed IFN-λ, receptor genes, But breast carcinoma231, MCF-7cells and malignant mesothelioma H2452, H226,211H, H28cells did not express IFN-λ, receptor genes. These data indicated that the expression of IFN-λ, receptor has its specificity among different cells.2IFN-λ mediated growth suppressionWe examined growth suppression of IFN-λ, in breast carcinoma cells, esophageal carcinoma cells, normal esophagus cells and malignant mesothelioma cells. The cells was treated with IFN-λ, and examined with the method of MTT. Among these cells, T.Tn cells shows IFN-λ-mediated growth suppression (P<0.05compared with untreated cells) in a dose-dependent manner; breast carcinoma231,453MCF-7cells, esophageal carcinoma TE1, TE2, TE10, YES6cells, malignant mesothelioma H2452, H2052, H226,211H, H28cells, normal esophagus Het-IA cells did not show IFN-λ-mediated growth suppression (P>0.05compared with untreated cells). These data indicate that IFN-λ, induced growth suppression is dependent on the cell types.3Combination of IFN-λ and anti-cancer agentsWe investigated possible combinatory anti-tumor effects produced by IFN-λ and representative anti-cancer agents for esophageal carcinoma T.Tn cells,5-FU and CDDP. esophageal T.Tn cells were cultured with5-FU (10uM) or CDDP (3uM).3.1We found that IFN-λ (lOng/ml) enhanced the cytotoxicity of5-FU (P <0.05). The data imply that there has a synergistic inhibition effect.3.2We found that IFN-λ(lOng/ml) enhanced the cytotoxicity of CDDP (P<0.05). The data imply that there has a synergistic inhibition effect.4Apoptosis induction with IFN-λ treatment4.1We investigated activated apoptotic pathways by examining cleaved caspase3in IFN-λ-treated cells, the respective cleaved forms was analyzed with Western blot. The data indicated that IFN-λ, can induced cells apoptosis.4.2We investigated activated apoptotic pathways by examining cleaved PARP in IFN-λ-treated cells, the respective cleaved forms was analyzed with Western blot. The data indicated that IFN-λ can induced cells apoptosis.4.3We investigated activated apoptotic pathways by examining cleaved caspase8in IFN-λ-treated cells, the respective cleaved forms was analyzed with Western blot. The data indicated that IFN-λ, can induced cells apoptosis.Conclusions:1The expression of IFN-λ receptor has restricted manner depend on the tissue or the cell line among a certain type of cancer.2IFN-λ induced growth suppression in some cell lines of certain type of cancer; Combination of IFN-λ, and chemotherapeutic agent (CDDP and or5-FU) produce a synergistic inhibition effect.3With the treatment of IFN-λ, the cleaved forms of caspase3、PARP、 caspase8increased, IFN-λ, induce cells apoptosis.