ABSTRACT
BACKGROUND: Multiple Myeloma (MM) is a heterogeneous, hematological neoplasm that accounts 2% of all cancers. Although, autologous stem cell transplantation and chemotherapy are currently the most effective therapy, it carries a notable hazards, in addition for being non curative. Recently, the Clustered Regular Interspaced Short Palindromic Repeats (CRISPR-cas9) has been successfully tried at the experimental level, for the treatment of several hematological malignancies. OBJECTIVES: We aimed to investigate the in-vitro effect of CRISPR-cas9-mediated knock-out of V-set pre B-cell surrogate light chain 1"VPREB1" gene on the malignant proliferation of primary cultured myeloma cells. METHODS: Bioinformatics' analysis was performed to explore the gene expression profile of MM, and the VPREB1 gene was selected as a target gene for this study. We knocked-out the VPREB1 gene in primary cultured myeloma cells using CRISPR-cas9, the VPREB1 gene editing efficacy was verified by determining VPREB1 gene expression at both the mRNA and protein levels by qPCR and immunofluorescence, respectively. Furthermore, the cytotoxic effect on primary myeloma cells proliferation was evaluated using cytotoxicity assay. RESULTS: There was a statistically significant reduction of both VPREB1 mRNA and protein expression levels (p<0.01). knock-out of VPREB1 gene in myeloma cell line resulted in a statistically significant reduction of myeloma cell proliferation. CONCLUSION: CRISPR-cas9-mediated knock-out of VPREB1 gene is effective for inhibiting the proliferation of primary myeloma cells. This would provide a basis for a promising therapeutic strategy for patients with multiple myeloma.
Subject(s)
CRISPR-Cas Systems , Immunoglobulin Light Chains, Surrogate/genetics , Multiple Myeloma/genetics , Cell Proliferation , Gene Editing , Genetic Therapy , Humans , Multiple Myeloma/pathology , Multiple Myeloma/therapy , RNA, Messenger/genetics , Tumor Cells, CulturedABSTRACT
BACKGROUND: Ovarian cancer represents an important problem in gynecologic oncology. A growing tumor induces the host endothelial cells to proliferate and supply the requisite vascular support allowing tumor development. Vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) have been demonstrated to induce angiogenesis in epithelial tumors in vivo. PATIENTS AND METHODS: This study included 24 tumors from patients with epithelial ovarian cancer in different stages, in addition to 20 tissue samples of benign ovarian lesions as a control group. VEGF has been measured in the cytosolic fractions using enzyme immunoassay and confirmed by Western blot analysis. Tissue IL-8 mRNA was assessed using reverse transcriptase polymerase chain reaction and immunohistochemistry for its protein. RESULTS: VEGF mean rank was significantly higher in ovarian cancer tumors compared to benign lesions (P < 0.001). Moreover, it was increased with advanced stages (P < 0.05) and in patients with poor survival (P < 0.05). Eight samples were positive for IL-8 mRNA, seven of them were in malignant group, with highest frequency in stages III and IV of the disease (6/12, 50%). IL-8 correlated with poor survival of the patients (P < 0.05). Log rank of Kaplan-Meier survival analysis was significant for FIGO stage, VEGF, and IL-8 (P < 0.05). CONCLUSION: These results indicate that VEGF and IL-8 are related to the malignant transformation process and can be considered as indicators of poor prognosis in epithelial ovarian cancer patients.
