Research On Target Therapy And Prognosis In Malignant Tumor

This study mainly researched on target therapy and pognosis in the non-Hodgkin lymphoma (NHL) and colorectal cancer (CRC) which are two common types of malignant tumor in China. The contents are divided into two parts as followed:Part1. Research on biomarkers associated with target therapy and prognosis in non-Hodgkin lymphomaIn this study, we evaluated the therapeutic efficiency of a new type of CRM1inhibitors with KPT-185and KPT-276in NHL in vitro and in vivo, the preclinical results reported here were expected to support the future clinical trials of these inhibitors in NHL. Furthermore, we tested some markers in peripheral blood of patients with DLBCL and evaluated the predictive value to identify the new indicators for DLBCL prognosis. The specific research contents as followed:Chapter1. Novel selective inhibitors of nuclear export (SINE) CRM1antagonists KPT-185for therapy in NHL. Here, we investigated the anti-tumor activity of KPT-185in eight NHL cell lines. The results showed that KPT-185displayed potent anti-proliferative properties at submicromolar concentrations (IC50values:60-120nM), induced cell-cycle arrest and apoptosis in NHL cell lines as well as patient cells. And the mechanism of anti-tumor effects mediated by KPT-185mainly involved in directly inducing the caspase families activity and downregulating the antiapoptoic proteins expression, such as CRM1, Survivin and NF-κB.Chapter2. The in vivo anti-tumor activity of novel CRM1inhibitor KPT-276in lymphoma. In this work, we characterized the anti-tumor effects of KPT-276in a xenograft NHL mouse model. Our results showed that tumor growth was significantly inhibited by orally administered KPT-276in all of the treatment groups, compared with vehicle control. Importantly, KPT-276exhibited the antitumor activity on even large xenografts. Besides, the mouse in100mg/kg treatment group was well-tolerated with no severely side effects observed. Histological analyses of xenograft tissue from KPT-276 treated animals showed the enhanced cancer cell nuclei staining of Survivin, which might be a potential molecular mechanism of KPT-276anti-tumor activity.Chapter3. Screening the new prognostic markers in peripheral blood of patients with diffuse large B cell lymphoma. In this study, the association of pretreatment of serum free light chain (FLC), serum total light chain (TLC), peripheral blood absolute monocyte count (AMC) and absolute lymphocyte count (ALC) with progression-free survival (PFS) and overall survival (OS) in diffuse large B cell lymphoma (DLBCL) were evaluated in103patients. The result showed that increased serum FLC κ and λ light chain or abnormal κ:λ FLC ratio was an independent, adverse prognostic factor for OS in DLBCL (P<0.01). Furthermore, elevated serum TLC κ and λ light chain was correlated with inferior OS and PFS (P<0.05). In addition, multivariate analyses demonstrated that low ALC was significantly associated with patient poor survival and was independent of the IPI (P<0.01).Part2. Research on molecular targets testing in colorectal cancerThis part mainly aimed to investigate the mutation status of KRAS and BRAF genes which are used as predictive markers of resistance to target therapy for patients with CRC. In this study, we initially compared two different assays for KRAS mutation testing to confirm the clinical applicability of direct sequencing. And then we investigated KRAS and BRAF mutations profile in a large scale of Chinese CRC populations. The specific research contents as followed:Chapter1. Comparative screening of KRAS mutations in colorectal cancer using direct sequencing and KRAS mutation analysis kit. This study detected KRAS gene mutations from66cases with colorectal cancer using direct sequencing and Applied Biosystems KRAS Mutation Analysis kit and assessed the sensitivity, the specificity and the accuracy of the two methods. Our data showed that the concordance between two methods reached95.5%(Kappa=0.907, P<0.001). And the direct sequencing method costed much less than the kit. Therefore, this method was proved to have the widespread clinical application value in assessing KRAS gene status for colorectal cancer patients.Chapter2. KRAS and BRAF mutation profile in Chinese colorectal cancer patients. In this work, direct sequencing was used to detect mutations in KRAS (codons12,13and61) and BRAF (exon11and15) in674Chinese CRC patients. The results showed that the positive rate of KRAS mutations was34.4%, including25.7%in codon12,6.8%in codon13, and2.1%in codon61. However, BRAF mutations were only detected in3.1%of the tumors, including1.9%in exon15and1.2%in exon11. KRAS mutation was more frequent in females than males (39.8%vs.30.9%, P=0.020), and patients older than60years exibited higher rates of mutation (39.0%vs.29.9%, P=0.015). But neither KRAS nor BRAF mutation correlated with smoking or drinking history, Dukes’ stage, histological type, tumor site, differentiation, lymph node metastasis, or distant metastasis.Taken together, we demonstrated the preclinical activity of the novel CRM1inhibitors in NHL in vitro and in vivo and evaluated the prognostic value of four markers in peripheral blood of patients with DLBCL. Furthermore, our data also cleared the wide application value of direct sequencing method for KRAS mutation testing and demonstrated the KRAS and BRAF mutation profile in Chinese CRC patients. These findings may provide a theoretical basis for the target therapy and prognosis in NHL and CRC.