| Ovarian cancer is one of the common malignant tumors in female genital organs, and its mortality rate ranks the first in gynecological tumors. It is estimated that about 15 per 100,000 people are diagnosed with new cases of ovarian cancer every year. Over the past 20 years, the 5-year survival rate of ovarian cancer has been hovering at a lower level, the recurrence rate within five years is as high as 80%, and recurrent time mainly concentrated in the treatment period of 3 years.Metastasis-associatedin colon cancer-1(MACC1) gene is the key regulation factor in the signaling pathway of hepatocyte growth factor / C-metprotoncogene(HGF/C-met), which can promote colon cancer cell proliferation, adhesion, invasion, and prompted proliferation and metastasis of colon cancer cells that induced by the hepatocyte growth factor. Mutiple studies have shown that in gastric cancer, liver cancer, pancreatic cancer and other malignant tumor tissues high expression of HGF and C- met can be detected, its over expression is associated with the recurrence and poor prognosis of malignant tumors. The main treatment of patients with ovarian cancer is surgery combined with chemotherapy. Recently, with the in-depth exploration of cilnical work and scientific research, treatment for ovarian cancer have been adjusted correspondingly, though treatment efficacy and prognosis of ovarian cancer patients didn't improve. Platinum compound was proved to be effective chemotherapy for the treatment of ovarian cancer, but patients with ovarian cancer 5 years of survival rate is only about 50% because of the existence of chemotherapy resistance.At present, there are many researches focus on the pathogenesis of ovarian cancer, but the relevant mechanisms of the molecular biology during the development of epithelial ovarian cancer is still not sure. At the same time, chemotherapy resistance is the main restriction factor for better prognosis to ovarian cancer. Therefore, there will be a wide prospect of clinical application in the drug resistance mechanism study of ovarian cancer chemotherapy, such as looking for the key molecular targets, developing high efficiency and low toxicity drug resistance reversal agent for the treatment of ovarian cancer, and so on.Objective To study the influence factors to the prognosis of patients with ovarian cancer, we analyzed clinical data, pathological character and survival time of patients with ovarian cancer in the first affiliated hospital of Zhengzhou University. Based on the preliminary study of the relationship between MACC1 and ovarian cancer which carried out by our research team, we deeply investigated MACC1, HGF, C-met and HE4 protein expression in ovarian cancer, discussed the relevance in the development of ovarian cancer, and provided clues for judging prognosis. By studying the varity of epithelial ovarian cancer cell proliferation and cancer cells cisplatin resistant sensitivity after the MACC1 gene interference, we were to explore the role of MACC1 gene in ovarian cancer chemotherapy drug resistance. Through the research of reversing ovarian cancer chemotherapy resistance by MACC1 gene in ovarian cell cells, we provided new theoretical basis for reversing ovarian cancer chemotherapy resistance and improving chemotherapy sensitivity clinically. Furthermore, we constructed transplant mouse model of ovarian cancer to explore the role of MACC1 in ovarian tumor proliferation..Methods 1. The Pearson chi-square, Kaplan Meier survival curve, log rank test and Cox regression were used to analyze the features of epithelial ovarian and prognosis of postoperative and point out the prognostic risk factors and independent factors which affect the prognosis. 2. The tissue chip, immunohistochemical staining were used to analyze the expression rate of MACC1, C- met, HGF and HE4 in cancer tissue and para-carcinoma tissue. 3. The Pearson chi-square, Kaplan Meier survival curve, log rank test and Cox regression are used to analyze features of epithelial ovarian cancer and prognosiswith different expression percentage. 4. Using cell culture technology cultivate resistant and sensitive cell line, RT-PCR and Western blot were used to test MACC1 m RNA and protein expression with different groups. MTT method was used to evaluate the change of different categories of cisplatin chemotherapy sensitivity with different groups. 5. Flow cytometry was used to test cisplatin dosing apoptosis with different groups. RT-PCR and Western blot were used to test the situation changes of ERK1/2, p-ERK1/2, caspase-3 and cleaved caspase-3 proteins and the effect of protein expression level with joining the U0126(ERK pathway inhibitor). 6. Cell culture technique was used to cultivate different processing cell. RT-PCR and Western blot were used to test MACC1 m RNA and protein expression with different groups and the weight of transplantation tumor and tumor growth with different groups in nude mouse.Results 1. Lymph node metastasis, clinical stage, pathological differentiation, residual tumor diameter and chemotherapy were independent risk factors for epithelial ovarian cancer cases prognosis. The dangerous degree in descending order for poor prognosis of epithelial ovarian were lymph node metastasis, pathological differentiation, chemotherapy, clinical stage, residual tumor diameter. 2. The expression rate of MACC1, C- met, HGF and HE4 in ovarian cancer tissues was higher than that in para-carcinoma tissues. The differences of tumor size, lymph node metastasis, clinical stage, residual tumor diameter and histopathological and chemotherapy were of statistical significance. 3. The median survival time of ovarian cancer patients with high expression of MACC1, C- met, HGF and HE4 was lower than patients with low expression. The expression level between each pair of MACC1, C- met, HGF and HE4 had correlation and synergistic effect. 4. Compared with blank group and the empty plasmid group, MACC1 m RNA and protein level were lower in MACC1 gene silencing group. The cisplatin halfinhibition concentration in MACC1 gene silencing group was lower than that in blank group and the empty plasmid group. After p-super-EGFP-MACC1 sh RNA transfection, MACC1 gene expression was down-regulated, and its m RNA and protein level was lower than blank group and the empty plasmid group. 5. Under the action of cisplatin with same concentration, SKOV3/DDP cells inhibition rate in MACC1 silent group was higher than untransfected group and empty plasmid transfection group. And the cell proliferation rate in MACC1 silent group was lower than the other two groups. The protein expression of p-ERK1/2 and caspase-3 was lower than untransfected group and empty plasmid transfection group, but the cleaved caspase-3 protein expression was higher. 6. In pc DNA-MACC1 group, the expression of MACC1 m RNA and protein, the weight of the nude mouse transplantation tumor and tumor growth were higher than untreated group and the empty vector control.Conclusion 1. Abnormal MACC1 gene expression was closely related to the development of epithelial ovarian cancer development, the synergistic effect of MACC1, C- met, HGF and HE4 was related to the malignant transformation of ovarian cancer cells. 2. It showed a increased tendency of MACC1 gene expression in SKOV3/DDP cells, silencing MACC1 gene can reverse SKOV3/DDP cells' cisplatin resistance effectively. 3. MACC1 may participate in cisplatin sensitivity adjustment of ovarian cancer and regulate ovarian cancer proliferation, apoptosis, migration and cisplatin sensitivity through the downstream of the ERK pathway. 4. MACC1 can produce a variety of biological behavior changes in epithelial ovarian cancer development and may become a new type of molecular targets in treatment of epithelial ovarian cancer. |
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