The Clinical Verification Of Multi-index Combined Diagnosis Of Ovarian Cancer Liquid Suspension Chip

CHAPTER I IDENTIFICATION OF CCL18?CXCL1?C1D?FXR1?TM4SF1?TIZ GENES/PROTEINS FOR THE DETECTION OF EARLY STAGE OVARIAN CANCER BY BIOINFORMATICS ANALYSISOBJECTIVE:Bioinformatics methods were used to discover and verify the intrinsic associations between CCL18? CXCL1? C1D? FXR1?TM4SF1 and TIZ genes/proteins,and their biological relevance to the early diagnosis of ovarian cancer.Constructing CCL18?CXCL1?C1D?FXR1?TM4SF1?TIZ PET-SUMO prokaryotic expression vectors,then producing and purifing high purity proteins.METHODS:(1)The gene/protein interaction networks of CCL18?CXCL1?C1D?FXR1?TM4SF1?TIZ based on the Gene MANIA online tool.Using Coremine medical online tools to annotate CCL18?CXCL1?C1D?FXR1?TM4SF1?TIZ biological processes.(2)CCL18?CXCL1?C1D?FXR1?TM4SF1 ? TIZ genes were connected with the PET-SUMO prokaryotic expression vector.The positive plasmids were transformed to express E.coli BL21 sensory cells.Under different conditions of IPTG to induce CCL18?CXCL1?C1D/ PET-SUMO plasmids expressed in E.coli BL21.Using the method of nickel affinity and refolding of urea-denatured/renatured to purify TM4SF1?FXR1?TIZ/PET-SUMO proteins.RESULTS:(1)Gene MANIA generated CCL18?CXCL1?C1D?FXR1?TM4SF1?TIZ genes/proteins interaction networks,and the 6 genes/proteins interacted with ovarian ovarian cancer and its early diagnosis.CCL18?CXCL1?C1D?FXR1?TM4SF1?TIZ genes/proteins are closely related to the biological processes have been marked out,showing that they have an associated with early diagnosis of ovarian cancer.(2)The CCL18?CXCL1?C1D?FXR1?TM4SF1?TIZ genes were successfully connected to the prokaryotic expression vector PET-SUMO,and the sequencing was consistent with the corresponding sequence in Genbank.(3)Under different conditions of IPTG to induce C1D?CCL18?CXCL1?TM4SF1?FXR1?TIZ/PET-SUMO plasmids to express in E.coli BL21.Resin soluble proteins C1D?CCL18?CXCL1/PET-SUMO were purified by nickel affinity.Using the method of the refolding of urea-denatured/renatured to purify inclusion body proteins TM4SF1?FXR1?TIZ/PET-SUMO.ALL proteins were verified by SDS-PAGE and Western-Blot.CONCLUSION:We successfully identified the intrinsic associations between CCL18? CXCL1?C1D?FXR1? TM4SF1? and TIZ genes/proteins,and their biological relevance to early diagnosis of ovarian cancer.We also successfully constructed CCL18 ? CXCL1 ? C1 D ? FXR1 ? TM4SF1 ?TIZ/PET-SUMO prokaryotic expression vector,and induced and purified the high purity proteins CCL18 ? CXCL1 ? C1 D ? FXR1 ? TM4SF1 ?TIZ/PET-SUMO.After SDS-PAGE and Western-Blot identification protein purification were the objective proteins and high purity.CHAPTER II CLINICAL EVALUATION OF THE MULTI-INDEX DETECTION OF OVARIAN CANCEROBJECTIVE: To construct a liquid suspension chip with multiple serum markers in patients with ovarian malignant tumor,and to measure the serum CCL18,CXCL1 antigen and C1 D,TM4SF1,FXR1,and TIZ autoantibodies and to verify the diagnostic value of liquid suspension microbubbles for ovarian malignant tumor.METHODS:(1)The recombinant fusion proteins of CCL18,CXCL1,C1 D,TM4SF1,FXR1 and TIZ were successfully purified by previous experiments.The carboxylated microspheres were coated with the corresponding antigens and antibodies to construct a liquid suspension chip with multi-index detection of serum in patients with ovarian malignant tumor.(2)Using liquid suspension microarray to detect 320 pelvic malignant tumor patients,148 benign pelvic tumor patients,100 healthy women and 100 female patients with liver cancer,100 female patients with breast cancer,,100 female patients with lung cancer and 100 patients with cervical cancer.(3)Establishing a combined test for the diagnosis of tumor and ovarian cancer diagnosis models combined with autoantibodies to diagnose the diagnosis of ovarian malignant tumor.Comparing the combined detection of six markers and CA125?HE4?CA19-9 alone in the diagnosis of early stage(I-II)ovarian malignant tumor.RESULTS:(1)This experiment constructed a stable liquid suspension chip detection system.(2)A diagnosis model of ovarian malignant tumor was constructed.CONCLUSION: The combined detection of six indicators is more effective than single CA125?HE4?CA19-9 in diagnosing ovarian malignant tumor.The combined detection of six markers was superior to single CA125?HE4?CA19-9 in the diagnosis of stage I and II ovarian malignant tumor.CHAPTER III PRELIMINARY EXPLORATION OF SERUM CCL18?CXCL1 ANTIGENS AND C1D? TM4SF1?FXR1? TIZ AUTOANTIBODIES IN JUDGING THE OUTCOME OF PLATINUM-BASED CHEMOTHERARY IN EPITHELIAL OVARIAN CANCEROBJECTIVE : To explore the effects of serum CCL18?CXCL1 antigens and C1D?TM4SF1?FXR1?and TIZ autoantibodies on the prognosis of outcome of platinum-based chemotherapy in epithelial ovarian cancer.METHODS: The serum CCL18,CXCL1 antigens and C1 D,FXR1,TM4SF1,and TIZ antibodies expression levels in patients with different clinicopathological factors were detected using a combination of multi-index diagnostic liquid suspension microarrays.The differences in clinical indicators of each index group were analyzed.The effect of index expression on the prognosis of epithelial ovarian cancer.RESULTS:(1)Analysis of the relationship between serum CCL18?CXCL1 antigens and C1 D ? FXR1 ? TM4SF1 ? TIZ antibodies expression and clinicopathological factors: various indicators of age,pathological type,FIGO stage,differentiation degree,postoperative residual size.There was no significant difference in the results of clinical pathological grouping patients(P>0.05);(2)Single factor analysis of prognostic factors in patients with epithelial ovarian cancer: Results found that OS and patient age,FIGO stage sooner or later,Postoperative residual size.(3)Multivariate analysis of prognostic factors in patients with epithelial ovarian cancer: age,type of pathology,and size of postoperative residual size are independent factors influencing patient prognosis.The expression levels of serum CCL18?CXCL1 antigens and C1D?FXR1?TM4SF1 and TIZ antibodies are not independent factors that affect the prognosis of patients.(4)The platinum-based chemosensitivity of patients with epithelial ovarian cancer is related to the FIGO stage ?the residual diameter of the surgery and age.Patients with a late FIGO staging?a postoperative residual diameter of 1 cm and the older one have increased risk of drug resistance.Serum CCL18,CXCL1 antigens and C1 D,FXR1,TM4SF1,and TIZ antibody expression levels were not significantly associated with risk of drug resistance.CONCLUSION: Serum CCL18,CXCL1 antigens and C1 D,FXR1,TM4SF1,and TIZ antibodies expression levels alone cannot be used to determine the outcome of a patient's chemotherapy.Simultaneous analysis of multiple markers may be a predictor of platinum-based chemotherapy outcome in patients with epithelial ovarian cancer.