Effect Of Pseudolarix Acid B On Human Endometrial Cancer Cell Ishikawa Apoptosis, Metastasis And Mechanism

Endometrial carcinoma is a reproductive tract disease which has become a serious threat to women’s health in our country. Endometrial carcinoma accounts for 20%～30% of female genital tract malignant tumor. The incidence rate of endometrial carcinoma is obviously increasing in the world. About 28% of patients have had distinct metastasis when diagnosis is made. Surgery is the main way to treat endometrial carcinoma, chemotherapy and radiotherapy are assisted methods. The toxicity of traditional chemotherapy drug is large, which is also harmful to the normal cells. It is still difficult to treat the late and recurrence of endometrial carcinoma. Therefore, it needs to find a kind chemotherapy drug of low side effects and inhibiting the distal metastasis tumors. Pseudolarix is the bark of the tree Pseudolarix kaempferi Gorden which is indigenous to central China. It is used in Chinese folk medicine to treat dermatologic fungal infections. Pseudolarix acid B (PAB) is one of biologically active components isolated from molecular Pseudolarix. PAB is a diterpenoid acid. Its molecular formula is C23H28O8. PAB has strong cytotoxicity. The main contents include:1. The study of apoptosis of human endometrial carcinoma cells Ishikawa induced by PAB. Endometrial carcinoma cell line Ishikawa was cultured conventionally, and observe the morphology of cell. Cell proliferation of Ishikawa treated with different concentration and application time of PAB and IC50 was detected by MTT method; Cell cycle and apoptosis affected by different concentration and application time of PAB in vitro were detected by flow cytometry;2. The study of the anti-metastasis effect of PAB to endometrial carcinoma cells Ishikawa. Cell clonality ability of Ishikawa treated with different concentration of PAB was assessed by clone formation assay; Cell wound healing ability after treated with different concentration of PAB was detected by wound healing assay; Cell invasion and migration ability after treated with different concentration of PAB was detected by transwell invasion assay;3. The mechanism of apoptosis and the anti-metastasis effect of human endometrial carcinoma cells Ishikawa induced by PAB. According to the cytology experimental results, Western-blotting was performed to detect the expression of signal pathway protein Akt, p-Akt, ERK1/2, p-ERK1/2 and the metastasis related protein VEGF, MMP9, E-cad to investigate the role of PAB play in endometrial carcinoma cells Ishikawa.Results:1. PAB inhibited the cell proliferation of endometrial carcinoma cells Ishikawa. IC50 of PAB to Ishikawa is 19.332 μM. PAB induced apoptosis of endometrial carcinoma cells Ishikawa, cell cycle arrested in G2/M phase.2. The migration and invasion ability of Ishikawa cells decreased significantly treated by different concentration and time of PAB. Cell clonality ability was decreased significantly after treated by PAB.3. The ratio of p-ERK1/2 in total ERK1/2 reduced, ratio of p-Akt in total Akt reduced after PAB treated. The expression of VEGF and MMP-9 decreased, with c-Myc and E-cad increased.Conclusion:1 Pseudolaric acid B can induce apoptosis of human endometrial carcinoma Ishikawa cells;2 Pseudolaric acid B can inhibit the metastasis of human endometrial carcinoma Ishikawa cells.3. Pseudolaric acid B can play its role though effecting the expressions of the key molecules MAPK in cellular proliferation signaling pathway and metastasis related genes such as MMP9,E-cad and VEGF.