| Cholangiocarcinoma is derived from the intrahepatic or extrahepatic bile duct epithelial tumors. In the liver primary tumors, the incidence of cholangiocarcinoma is lower than the liver cancer. There is a high degree of malignancy with occult its incidence, clinical symptoms and signs appeared later. which accounts for less 2% of all human malignancies, accounting for 3% of gastrointestinal cancer. In 1840, Durand Fardel firstly proposed cholangiocarcinoma, In recent years, the incidence and mortality of cholangiocarcinoma upward trend, the survival of 5-year is less than 5%. More tend to comprehensive treatment based on surgery, even only part of the early surgical treatment for patients. Since the special position of cholangiocarcinoma, the bile duct tumors have not been completely blocked in patients with subclinical before. So it is difficult to make an early diagnosis. Most of the clinical patients are treated in end of stage. Because of the advanced or metastatic of the tumors, about 80% of cholangiocarcinoma patients are only treated with palliative treatment. The conventional methods of cholangiocarcinoma are not sensitive to radiotherapy and chemotherapy. It is great significance to explore a new and effective therapeutic approach to improve its prognosis.Monomer bufalin, the traditional Chinese medicine, originates from the Chinese big toad and the black frame toad ear the gland and the skin gland secretion's size, the molecular formula is C24 H34 O4, relative molecular mass is 386.5. A great many of researches have indicated that this ingredient has lots of medical functions, such as anticancer treatment, cardiotonic effect, toxicity and anaesthesia. The machines of induction tumour cell differentiation and perishe are possible to suppresses the leukemia cell's multiplication through the disturbance of mitotic cycle, the changing of perishes in related gene, such as bcl-2 family, inhibit the abnormal activation of MAPK and so on. There have been a lot of studies reported in liver cancer, stomach cancer, the colon cancer, however, only few of reports have concerned cell's function and the machine-made aspect of bile duct cancer. We use the bufalin monomer function in human bile duct cancer cell, named QBC939, to examine the suppressing of cyclin E, P27 mRNA and the protein expression. The purpose of this stuy is to explore the influence of bufalin on the bile duct cancer cell, its possible molecular mechanism and then provide the theory basis for the clinical therapy.ObjectiveThe aim of this study is to observe the effect of bufalin on the expression of P27 and Cyclin E in cholangiocarcinoma cell line QBC939, investigate the pathways of bufalin in inhibiting tumor cell proliferation, and explore its antitumor mechanism.Methods1. Cholangiocarcinoma cell line QBC939 was cultured by routine method, and then treated with different concentrations of bufalin (0.1μM, 1μM, 10μM).The proliferation apoptosis and cell cycle of QBC939 cells were investigated by MTT and Flow cytometry.2. Bufalin toxicity was assayed by Lactate dehydrogenase (LDH).3. RT-PCR and Western blot were performed to measure the expression of P27 and Cyclin E at gene and protein level.Results1. MTT assay results showed that of bufalin on the growth of human cholangiocarcinoma cell QBC939 inhibited, and in a certain range of time-and concentration-dependent. 2. Bufalin does not through its toxic effects to inhibition of cell proliferation QBC939.3. With different concentrations of bufalin for 24 hours, cholangiocarcinoma cell line QBC939 cell cycle arrest in G0/G1, with a good dose-effect relationship.4. By RT-PCR and Western blot assay, we found that bufalin at different concentrations for 24 hours, in cholangiocarcinoma cell line QBC939 decreased expression of cyclin E, p27 expression increased.ConclusionBufalin may inhibit the proliferation of QBC939 cells by down-regulating the expression of Cyclin E and up-regelating the expression of P27, blocking cell cycle.
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