| Background and designOnce discovered,70% of ovarian cancer is in the advanced stage. With a 25%-30% five year survival rate, the death rate of it tops gynecological malignant tumors. Therefore, once diagnosed with ovarian cancer, a good many patients suffer mind and mental stress and lose heart to therapy, which seriously affects the prognosis.For the past few years, as the medical model improves from a single biomedical model to a bio-psycho-social model, the scholars at home and abroad pay close attention to the effect of the psychosocial factor on tumor patients. However, its action mechanism is not clear. Now mental intervention therapy and immunotherapy are two important contents of tumor combined therapy. But no unified opinion has reached on the therapeutic efficacy. Thus, a deep study on the relation between psychology and tumor development, prevention of tumor occurrence and growth by means of psychological factor and psychotherapy, tumor control by altering patients'immune state to raise survival rate and improve life quality, all these remains to a further research. Since psychological factor is hard to control, stress animal model is adopted to study the effect of psychological factor on tumor, which bears advantages on the aspects of standard, reproducibility, steerability, short observation cycle and ideal results. This study concerns two aspects. On one hand, by setting up bearing-cancer nude mice dys-psychological stress model, it investigates the relation between dys-psychological stress and ovarian tumor immunity, and the effect of dys-psychological stress influence on the tumor's growth in bearing-cancer nude mice.On the other hand, it focuses on both gynecological malignant tumor patients'social psychology features and mind-physique health status by using the clinical psycho- rating scale and questionnaire.The ultimate purpose is to provide theoretical foundation for psychotherapy and immunotherapy to ovarian cancer. Part I Experimental research on dys-psychological stress effect on human epithelial ovarian carcinomaObjective:To establish dys-psychological stress model using nude mice bearing human epithelium ovarian carcinoma. Giving therapy with DDP, observing dys-psychological stress effect on the growth of subcutaneous xenotransplanted tumor in nude mice bearing human epithelium ovarian carcinoma,and the influence on p53 and NFκBp65 expressions,and neuroendocrine and immune function changes as well. Discussing the action of dys-psychological stress influence on the tumor in bearing-cancer nude mice. The purpose is to provide theoretical foundation for psychotherapy and immunotherapy to ovarian cancer.Methods:1. Establishing a subcutaneous xenotransplanted tumor model with human epithelium ovarian carcinoma in nude mice and to do tumor passage transplantation in nude mice .When the tumor under observation reached the size of 0.8cm-1cm in diameter, it was cut off and into pieces of 1mm-2mm in diameter,then it was transplanted subcutaneously into another nude mice. Examined all the tumor pathologically. And made the tumor mass into single cell suspension to culture it in vitro and observe chromosome features. This research adopted the fourth-generation human epithelium ovarian cancer tissue.2. Setting up dys-psychological stress model with restraint, putting each nude mouse into a 50ml centrifuge tubes ( with 3mm holes, excellent ventilation), not squeezing their tails, enabling them to move to and fro in the tubes. Freeing the mice after 8h and resuming food and water. Restraining the mice 8h/day for two weeks (10:00AM-6:00PM) stress 5d/week, and preventing adaptation. The second day after the experiment, blood samples were drawn from orbit, kill the nude mice, cut and weigh the tumors and examine them.3. Dividing the mice into six groups(1)normal group (negative control group): NS injection introperitoneally;(2)normal group with stress ( positive control group ) : NS injection introperitoneally and with restraint; (3)tumor group (negative tumor control group): NS injection introperitoneally;(4) tumor therapy group (positive tumor control group): DDP injection introperitoneally and with restraint;(5) tumor with stress group (experiment group): NS injection introperitoneally and with restraint;(6)tumor therapy with stress group (experiment group): DDP injection introperitoneally and with restraint.Group (1) and(2)have no tumor; group(3)to(4)have tumor. The NS dose for group(1), (2) ,(3) and (5) is 0.2ml/20g , and the DDP dose for Group (4) and (6) is 3mg/kg, 0.2ml/20g ( Preparing DDP into the injection with a 0.3mg/ml concentration.), injecting introperitoneally once 3 days, totaling 4 times.4. Observing the nude mice's living state, tumor growth ( tumor size and weight, repress rate) and histological and morphological changes.5. Examining the expressions of p53 and NFκBp65 in each group's tumor tissue by means of Western Blotting.6. Examining corticosterone, 5-hydroxytryptamine , norepinephrine , IL-2, IL-6 and TNF-αchanges in each group's nude mice serum by means of ELISA. Examining NK cell activity in each group's nude mice spleen tissue by means of MTT.Results:1. A subcutaneous xenotransplanted model of human ovarian cancer was established successfully in nude mice. The success rate of tumor passage transplantation in nude mice is 100%. Detect the tumor metastasis in nude mice's livers and spleens. Histological observation showed similar morphological features to human serous epithelium ovarian cancer, which accords with low differentiation of serous adenocarcinoma and bears human chromosome features. Based on this,a dys-psychological stress model was established successfully in nude mice bearing human ovarian cancer with restraint.2. Comparison of nude mice weight and tumor weight.(1) The dys-psychological stress groups were in a bad state of growth, feeding and action. The tumor with stress group was the worst. (2) The mice in restraint groups weighed less than their counterparts in non-restraint groups, differing significantly ( P﹤0.05 ) , showing that the dys-psychological stress made the mice lose weight significantly. The tumor therapy group weighed less than tumor group(P﹤0.05), showing that chemotherapy made the mice lose weight significantly.(3) Tumors in each restraint groups weighed more than the corresponding non-restraint groups(P﹤0.05).3. Comparison of the expressions of p53 and NFκBp65(1) The expressions of p53 and NFκBp65 in each restraint group were enhanced compared with the corresponding non-restraint group ((P<0.05).(2) The expressions of p53 and NFκBp65 in tumor group were stronger than those in tumor therapy group (P<0.05).(3) Comparing the groups from 3) to 6), p53 and NFκBp65 expressed the strongest in tumor with stress group, coming next is tumor therapy with stress group, followed by tumor group, and tumor therapy group was the weakest.4. Comparison of CORT,NE and 5-HT values in nude mice serum(1)The serum corticosterone and NE levels in each stress group were higher than those in the corresponding non-stress groups(P﹤0.05); the serum 5-HT levels in normal group were higher than that of restraint group but with no significant differences(P﹥0.05);. and that in tumor with restraint group were higher than that in tumor group but with no significant difference. The serum 5-HT levels in DDP with restraint group were lower than those in tumor therapy group(P﹤0.05).(2) CORT,NE and 5-HT levels in tumor groups were higher than those in normal groups (P﹤0.05).(3) CORT,NE and 5-HT levels in tumor therapy group is higher than those in tumor group(P﹤0.05).(4)The NE levels in tumor therapy group were higher than those in tumor group(P﹤0.05). The CORT and 5-HT levels in tumor therapy group were lower than those in tumor group(P﹤0.05).5. Comparison of TNF-α,IL-2 and IL-6 values in nude mice serum and NK activity values. (1)The serum TNF-αand IL-6 levels in each restraint group were higher than those in the corresponding non-restraint groups(P﹤0.05); the serum IL-2 levels in each restraint group were lower than those in the corresponding non-restraint groups(P﹤0.05).(2)The serum TNF-αand IL-6 levels in tumor group were higher than those in normal group;IL-2 and NK activity levels were lower (P﹤0.05).(3)TNF-α,IL-6 levels and NK activity levels in tumor therapy group were lower than those in tumor group(P﹤0.05). IL-2 levels were lower than those in tumor group,but with no significant differences(P﹥0.05)(4)The serum TNF-αand IL-6 levels in tumor stress group were higher than those in normal stress group;IL-2 and NK activity levels were lower (P﹤0.05).Conclusion:1. Setting up dys-psychological stress model with restraint successfully.2. The dys-psychological stress adds the tumor weight significantly. Chem- otherapy can restrain tumor growth, but with stress, it can accelerate the growth, resulting in tumor weight increase.3. Under both chemotherapy and non-chemotherapy conditions, dys- psycholo- gical stress enhances P53 and NFκBp65 protein expressions in the subcutaneous enotransplanted tumor. The DDP chemotherapy weakens the two expressions, showing chemotherapy effect,but added with restraint stress, it makes the expressions stronger than those before the chemotherapy. It shows that by enhancing P53 and NFκBp65 protein expressions,the chemotherapy effects can be weakened, thus promoting tumor growth.4. Activating hypothalamus-pituitary-adrenal axis, dys-psychological stress causes neuroendocrine dysfunctions,hence CORT and NE excessive synthesis, and 5-HT disorder. And with intervention of CNS,they can disturb the normal function of brain psycho-adjusting, weakening immunity function. Under chemotherapy state, dys-psychological stress makes the neuroendocrine dysfunction more serious.5. Dys-psychological stress significantly increases TNF-αlevel and IL-6 level and lowers NK activity and IL-2 level, proving that dys-psychological stress can influence immunity function of the nude mice bearing human tumor. DDP chemotherapy significantly increases TNF-αlevel, IL-6 level and decreases NK activity, proving that DDP chemotherapy can decrease immunity function of the nude mice bearing human tumor. Especially in the state of chemotherapy, dys-psychological stress weakens immunity function further.Part II Psychological status of patients with gynecological malignant tumors and correlation factor analysisObjective:To investigate psychological status of patients with gynecological malignant tumors and mind-physique health status.Methods:Clinical psycho-rating scale: life event scale (LES) and symptom checklist-90 (SCL-90), is applied to test life events and mind and physique health status of 60 cases of patients with gynecological malignant tumor. The results were compared with those of 40 female patients with benign tumors and 40 normal female respectively.Results:1. The frequency and tension value of total life events and those of negative life events in the malignant cancer group were much higher than those in both the benign tumor group and normal group(p < 0.05)and no remarkable changes in positive life events were detected. The frequencies of total life events and negative life events were higher in the benign tumor group than those in the normal group (P﹤0.05). There were no significant changes in the frequency of positive life events, total tension value, and positive tension value in both groups.2. Each index of SCL–90 in the malignant cancer group,except the average positive scores and other factor scores showing no significant difference, was higher than that in the benign tumor group and the normal group(P﹤0.05).Except that the average positive scores and the depression scores were higher than those in the normal group (P﹤0.05), other indexes in the benign tumor group showed no significant difference. Conclusion:Dys-psychosocial factors are closely linked to the occurrence and growth of gynecological malignant tumors and active psychological intervention should be performed on the patients. |
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