Chapter 2:Study On Expression Levels Of ITGB6,PIK3R3,NCALD,LAMA3 And Clinical Outcome In Platinum-resistant Ovarian Cancer

Objective: To confirm methylation level of ITGB6,PIK3R3,NCALD and LAMA3 in Platinum-resistant ovarian cancer,explore the relationship between methylation and clinical outcome.Methods: The methylation status of ITGB6,PIK3R3,NCALD and LAMA3 were detected by pyrosequencing.Results:(1)The methylation level of ITGB6 、 NCALD 、 LAMA3 was higher in the Platinum-resistant ovarian cancer;There was no significant difference in PIK3R3 methylation between the resistant group and the sensitive group.(2)The OS was shorter in ITGB6、NCALD 、LAMA3、PIK3R3 hypermethylation patients.Conclusion: The methylation level of ITGB6,NCALD and LAMA3 is an effective biomarkers to predict chemoresistance and prognosis in ovarian cancer.Objective: To confirm expression level of ITGB6,PIK3R3,NCALD and LAMA3 in Platinum-resistant ovarian cancer,explore the relationship between ITGB6,PIK3R3,NCALD and LAMA3 expression and clinical outcome,investigate the relationship between expression and methylation.Methods: The expression status of ITGB6,PIK3R3,NCALD and LAMA3 were detected by QRT-PCR and immunohistochemistry.Results:(1)The expression of NCALD was down in the drug resistant group;The methylation level of NCALD(chr8:102804678,chr8:102804702)was negatively correlated with m RNA expression;The PFS was longer in the NCALD high expression group.(2)There was no significant difference in PIK3R3 expression between the resistant group and the sensitive group;The PFS was shorter in the NCALD high expression group.(3)The expression of LAMA3 was down in the drug resistant group;The methylation level of LAMA3(chr18:21452788,chr18:21452819)was negatively correlated with m RNA expression;The PFS,OS was longer in the LAMA3 high expression group.(4)There was no significant difference in ITGB6 expression between the resistant group and the sensitive group.(5)The sensitivity and specificity of combined expression in the diagnosis with chemoresistance were0.692 and 0.691,AUC=0.730.Conclusion:The expression levels of NCALD and LAMA3 is an effective biomarkers to predict chemoresistance and prognosis in ovarian cancer.NCALD and LAMA3 may be involved in drug resistance through regulation of gene expression by methylation.Objective: To detect the methylation level of ITGB6,PIK3R3,NCALD and LAMA3 in the ovarian cancer cells and nude mouse animal models.Methods: The methylation level of ITGB6,PIK3R3,NCALD and LAMA3 were detected by pyrosequencing.Results:(1)There was no significant difference in ITGB6 methylation between SKOV3 and SKOV3DDP;The methylation level of ITGB6 in cisplatin-resistant nude mouse was lower than cisplatin-sensitive nude mouse.(2)The methylation level of NCALD was higher in SKOV3DDP;The methylation level of NCALD in cisplatin-resistant nude mouse was higher than cisplatin-sensitive nude mouse.(3)The methylation level of LAMA3 was lower in SKOV3DDP;The methylation level of LAMA3 in cisplatin-resistant nude mouse was lower than cisplatin-sensitive nude mouse.(4)The methylation level of PIK3R3 was higher in SKOV3DDP;The methylation level of PIK3R3 in cisplatin-resistant nude mouse was higher than cisplatin-sensitive nude mouse.Conclusion: The methylation levels of PIK3R3 and NCALD were higher in drug-resistant cells and cisplatin-resistant nude mouse.