The Effect Of Herbicide Atrazine On Proliferation And Metastasis Of Human Ovarian Cancer Cell Skov3in Vitro

Background：atrazine (ATR)，a trazine herbicide, have been a mainstayof preemergent pesticides for a number of decades and with low human toxicity.ATR has the characteristics of the huge amount of use, the long residual periodand the wide range of pollution. ATR is frequently detected in ground andsurface water,and its persistence in the environment and bioaccum ulation mayconstitute a significant threat for human health.Our goverment set0.003mg/Las the minimum level of ATR in surface water..In the surface water in EastLiaohe River Basin in jilin, the average atrazine content in waters in regionswith glebe or without glebe were namely9.71μg/L and8.854μg/L in2005.ATR has the recognized effect not only on the health of the people who applythe pecticides, but also on the health of those consume the contaminatedproducts. Epidemiological studies on the Nextipac community in Jalisco,Mexico show that: there exist health hazards for those farm workers exposed to ATR, at organic and cellular levels. RinskyJL et al reviewed the datas onKentucky birth certificate for the years2004-2006and assessed existing datasources for atrazine levels in public drinking water for the years2000-2008,and found a positive association between atrazine and preterm birth. Theinternational researches about ATR focu on the nervous system toxicity, theimmune system toxicity, the endocrine system toxicity and promoting on theprogression of tumor. Ovarian cancer is a highly malignant tumor, with highMortality and low five-year survival rate (the mortality rate of serous ovariancancer was90%, and the five-year survival rate was13%). ATR is able to affectestrogen production by inducing aromatase, which converts androgen intoestrogen. The effect of long-term high level of estrogen can lead to ovariantissue malignant transformation and lead to the occurrence of ovarian cancereventually. The DNA damage induced by peroxidation of ATR is one of theimportant incentives of tumorigenesis. Whether ATR in environmentallyrelevant dose promoting on the progression of ovarian cancer or not? It has notbeen reported. We have exploited the affects of ATR on proliferation andmetastasis ability of ovarian cancer cell in environmentally relevant dose and investigated the mechanism underlying it. It can provide theoretical andexperimental basis for the correct use of ATR and the effective prevention oftumorigenic poison. It can also provide scientific datas for environmentalmonitoring of ATR and the prevention and treatment of ovarian cancer.Objective: To test the effect of ATR on proliferation and invasion ofovarian cancer cell in environmentally relevant dose in vitro.Method: Serous cystadenocarcinoma has the highest incidence in humanovarian cancer and We chose the serous cystadenocarcinoma ovarian cancercell lines Skov3as the experimental cell. Skov3cells were cultured in vitro andexposed to environmentally relevant doses of ATR (0.1μmol/l and1μmol/l).We used MTT and colony formation assay to test proliferation of Skov3cell,and used immunohistochemical staining to analyze the expression of PCNA.The changes in cell cycle progression were detected by flow cytometry. Toanalysis the molecular mechanism of cell cycle progression induced by ATR,PT-PCR and Western-blot were used to detect the changes on transcription andexpression levels of P21, P53and CyclinE. The change on invasion andmetastasis of Skov3cell were detected via scratch test and transwell invasion, espectively. We used Western-blot to detect the expression of MMP-2andMMP-9to analysis the mechanism of ovrian cancer invasion and metastasisability promoted by ATR. The changes on the expression of VEGF andSurvivin were detected to analysis the changes on the ability of Skov3cell toform new blood vessels via western-blot.Result: ATR in the doses of0.1μmol/l and1μmol/l can enhance Skov3ovrian cancer cell growth and colony formation. It can also up-regulate theexpression of PCNA in nucleus. ATR can promote cell to pass the G1/S phasecheckpoint, increase the ratio of cells in S phase. We found that ATR maybepromote cell cycle progression by repressing the transcription P53and P21,down-regulating the expression of them, up-regulating the expression of CyclinE and Survivin. ATR enhanced the invasion and metastasis ability of Skov3though up-regulating the expression of MMP-2, MMP-9and VEGF.Conclusion: ATR can promote the proliferation and invasion of ovariancancer Skov3cell. ATR may enhance the growth by regulating the expressionof P53, P21,Cyclin E and Survivin and promote the process of Skov3invasionby up-regulating the expression of MMP-2, MMP-9and VEGF. ATR may also improve the survival rate of ovarian cancer cell and promote the process ofneovascμlarization by up-regμlating the expression of Survivin and VEGF.