| Background: Ovarian cancer is one of the three major malignant tumors of gynecology,the incidence rate ranks the third China,the current ovarian cancer is the highest mortality in gynecological tumors of malignant tumors.The main cause of high mortality in ovarian cancer is that 70% of patients have been diagnosed in the late stage,these patients will have more than 70% of the recurrence of the tumor;and many patients with drug resistance and eventually lead to death.The In recent years,the relevant research put forward the view of ovarian cancer stem cells,suggesting that the presence of ovarian cancer stem cells may be one of the important factors in the recurrence of ovarian cancer,the presence of stem cells to enhance the tumor formation and drug resistance.So far,ovarian cancer stem cell research has been challenging,how to identify such a special biological behavior of the cell population to become the focus of the current stem cell research and difficult;so to find different tumor stem cell surface expression of different biomarkers become identified stem cells One of the important ways.At present,the study of ovarian cancer stem cell related markers are CD133,CD24,CD44,CD117 and ALDH1.Related literature reported,the current CD133,CD44 and CD117 has been widely considered to be a specific marker of ovarian cancer stem cells,but CD24 and ALDH1 are not definite conclusion.ALDH1 is also considered to be a marker of tumor stem cells in many solid tumors and is not yet known in ovarian cancer stem cells.However,the expression of ALDH1 is currently detected in some human ovarian cancer tissues and cell lines,In cells line,ALDH1-expressing cells are more resistant to chemicals,and ALDH1-expressing cells can promote tumor cell growth compared with ALDH1-negative cells.Therefore,ALDH1 can also be the same as CD133 ovarian cancer stem cells as a specific marker,has become the focus of the current study.At present,the experimental method of stem cells is used to induce the suspension of cell spheres with stem cell characteristics.Therefore,in this study,the suspension cells with stem cell characteristics were cultured by immunofluorescence.The expression of CD133 and ALDH1 in suspension cells were detected by immunofluorescence cases,compared with the cell line expression differences,which initially infer whether ALDH1 can become a marker of ovarian cancer stem cells.Objective: The suspension cell spheres were cultured by serum-free from cell lines and drug resistant cell lines.that is ovarian cancer stem cells.The expression of CD133 and ALDH1 in suspension cell and cell line and drug resistant cell line were detected.It was concluded that ALDH1 could be consistent with CD133 as ovarian cancer stem cells.Methods: We chosed the ovarian cancer cell lines SKOV3,Hey-A8 and OVCAR-8 to select the better cell lines.And established its drug-resistant cell lines Hey-A8-DDP,OVCAR-8-DDP.The expression of CD133 and ALDH1 in Hey-A8,OVCAR-8 and drug-resistant strains Hey-A8-DDP and OVCAR-8-DDP were detected by Western-blot.The ovarian cancer cell lines Hey-A8,OVCAR-8 and drug-resistant cell lines Hey-A8-DDP and OVCAR-8-DDP were cultured in serum-free culture.Immunofluorescence staining was used to detect the expression of CD133 and ALDH1 in cultured cell spheres of cell lines and drug resistant cell lines.Results: 1.the ovarian cancer cell lines Hey-A8 and OVCAR-8 and their drug-resistant cell lines Hey-A8-DDP and OVCAR-8-DDP were successful cultured cell spheres in serum-free medium.The cells were cultured on the 8 day,Compared with the cell lines Hey-A8 and OVCAR-8,,the cell spheres of their drug-resistant cell lines were larger,more regular and more spherical.2.the expression of ALDH1 in Hey-A8-DDP was higher than that in the ovariancancer cell line Hey-A8.There was no significant expression of ALDH1 in ovarian cancer cell line OVCAR-8 and drug resistant cell line OVCAR-8-DDP.No expression of CD133 was detected in these cell lines and drug resistant cell lines.3.ovarian cancer cell lines Hey-A8,OVCAR-8 and its drug-resistant cell lines Hey-A8-DDP,OVCAR-8-DDP cultured suspension cell spheres,CD133,ALDH1 were detected by Immunofluorescence staining,And the expression of both in the suspended cell spheres of drug resistant cell lines was higher.,And the both of changes in OVCAR-8 cell line and drug resistant cell line OVCAR-8-DDP were more obvious.Conclusion:1.the ovarian cancer stem cells were screened in the ovarian cancer cell lines Hey-A8 and OVCAR-8 and the drug-resistant cell lines Hey-A8-DDP and OVCAR-8-DDP,and the ovarian cancer stem cells are enriched in drug-resistent cell line,and thus infer the presence of ovarian cancer stem cells is one of the important reasons for the recurrence of ovarian cancer resistance.Conclusion: 1.the ovarian cancer stem cells were screened in the ovarian cancer cell lines Hey-A8 and OVCAR-8 and the drug-resistant cell lines Hey-A8-DDP and OVCAR-8-DDP,and the ovarian cancer stem cells are enriched in drug-resistent cell line,and thus infer the presence of ovarian cancer stem cells is one of the important reasons for the recurrence of ovarian cancer resistance.2.ALDH1 is highly expressed in cultured cell spheres that can represent ovarian cancer stem cells,and expressed or not expressed in the cell line and the resistant cell line.suggesting that ALDH1 may be the same as CD133,ovarian cancer stem cells become a meaningful biological Learn to mark.3.CD133,ALDH1 in all the cultured suspension of the ball in the common high expression,the two combined detection,can increase the success rate of ovarian cancer stem cell screening. |
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