| Malignant tumor remains one of the most major causes of mortality and a common dieases threating human health worldwide.Traditional treatments in clinical can hardly cure cancer fundamentally and accompany by significant side effects.In recent decades,with the in-depth study of tumor molecular mechanism uncreasingly,TBCT(targeted based cancer therapy),which targets specific molecule that play a key role for the occurrence of cancer and its regulatory molecules in signal transduction pathways arise a concept change in cancer treatment.TBCT as a novel therapeutic strategy has been applied successfully and effectively in clinical due to its high specificity,selectivity and low side-effect.The determination of accurate therapeutic target is both the key of TBCT and rmains the difficulties to conquer cancer.There are several kinds of monoclonal antibodies as cancer molecular targeted drugs had been applied in clinicalcation and clinical trials or on the market since these macromolecules showed good efficacy and had none or small adverse reactions ocuured.Successive discoveries suggested CD98,a TAM(tumor associated molecule)played an extensive and significant role due to its high levels in human diseased or normal tissues in recent years.The current study concentrated on exploring CD98 expression in different cancer tissues and its potential application in diverse clinical settings including risk assessment,detection and localization as a cancer-associated biomarker.CD98 had been reported that involved in biological events such as tumorigenesis,proliferation and abnormal metabolism.While all these malignant phenotypes can be reversed when CD98 is interfered,knocked out or even blocked.Therefore,our study focused on the screening for novel monoclonal anticancer antibodies targeting CD98 and its related molecular mechanism.This present work includes three parts as follows:Part ?: CD98,a potential diagnostic cancer-related biomarker,and its prognostic impact in lung cancer and colorectal cancer patients.On immunohistochemical analysis of 10 TMAs with 713 samples of carcinoma tissues and 260 samples of normal or cancer adjacent normal tissues,we found CD98 was expressed in carcinoma with an overall positivity rate of 74%,which was significantly higher than those in adjacent normal or normal tissues,and suggested that it might be a valuable biomaker.In addition,a retrospective study of pathological sections of 100 matched pairs of normal and tumor tissues from patients confirmed the high expression of CD98 is associated with poor prognosis of colon cancer and can be used as an independent prognostic factor in CRC.Part ? Screening of antibodies against CD98 based on the human antibody transgenic mice technologyFirst of all,we constructed a system of prokaryotic expression of CD98 antigen.Furtherly,we used the human antibody transgenic mice to screen stability high expression monoclonal antibody against CD98.Specifically,the human antibody transgenic mice was immuned by the expressed CD98,and then hybridoma technique was adoped to prepare mAbs against CD98.By the method,we obtained four cell lines which expressed human antibody against CD98.Part ? the identification and functional studies of the obtained mAbs against CD98In this part,we first identified the established cell strains by Western blot,immunohistochemistry assay.The results showed that four human CD98 antibodies CD98-1,CD98-3,CD98-4 and CD98-5 have a farely good affinity and specificity against the target antigen.In addition,the antibodies prepared could interact with prokaryotic CD98 by ELISA.The effect of purifed mAb CD98-1 could inhibit the proliferation of lung cancer cells and affect the growth of lung cancer cells detected by CCK8 test.In all,the preparation of human monoclonal antibodies against CD98 provides new options and strategies for tumor targeted therapy,and the related work also provides a basis for understanding the function and mechanism of the molecule in tumors and normal tissues. |
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