ABSTRACT
Objective: To observe the expression of valosin-containing protein CVCP) in the epithelial ovarian cancer (EOC) tissue and its relationships with the clinicopathological features of the EOC patients∗ and to provide the basis for the molecular treatment of EOC. Methods: The expressions of VCP in 94 EOC tissue samples, 13 ovarian borderline tumor tissue samples, 36 ovarian benign tumor tissue samples and 8 normal ovarian tissue samples were measured by immunohistochemical method. The relationships between the VCP expression and the clinicopathological parameters (age, FIGO stage, pathological type, histological grade, lymph node metastasis or not, ascites or not, preoperative CA125 level) of the EOC patients were analyzed. The expressions of VCP in human ovarian epithelial HOSEPIC cells and human EOC SKOV-3 cells were detected by immunofluorescence technique and Western blotting method. The SKOV-3 cells were divided into control group (without CB-5083) and treatment group (with CB-5083)∗ the migration rates of the SKOV-3 cells in two groups were analyzed by scratch experiment, and the clone formation rates of the SKOV-3 cells in two groups were analyzed by plate clone formation experiment. The expressions of VCP in the cells in two groups were analyzed by immunofluorescence technique. The expression levels of VCP, NF-kB/P65» IieBa∗ and p-Iiefta proteins in the SKOV-3 cells in two groups were detected by Western blotting method. Results: The positive expression rates of VCP in EOC, borderline ovarian tumor, benign ovarian tumor and normal ovarian tissues had significant difference ( P0. 05). The VCP expression level in the SKOV-3 cells was higher than that in HOSEpiC cells ( P<0. 05). The migration rate of the SKOV-3 cells in treatment group was lower than that in control group ( P<0.05), the cloning formation rate of the SKOV-3 cells in treatment group was lower than that in control group ( P<0. 05). The expression levels of VCP and NF-kB/P65 proteins of the SKOV-3 cells in treatment group were lower than those in control group (P'<0. 05) ∗ and the expression level of p-IieBa protein was higher than that in control group ( P< 0. 05). Conclusion: VCP is highly expressed in the EOC tissue and cells∗ and high-expression VCP can promot the tumor migration and proliferation.
ABSTRACT
Objective To investigate the radioresistance factors in non-small cell lung cancer (NSCLC)cell line A549, and provide new targets for radiotherapy sensitization drugs development. Methods Establish the stable model of radioresistant NSCLC cell line A549 under irradiation; investigate the whole-transcriptome alteration of radioresistance cell line and radiosensitive cell line using gene expression microarray; perform bioinformatic approaches gene ontology (GO) analysis and Pathway analysis. Results The expression profile microarray showed that 1410 differentially expressed genes (733 up-regulated and 677 down-regulated) were detected in resistant and sensitive strains; GO analysis showed that it was mainly related to cell cycle and DNA replication; Pathway significant enrichment analysis showed that mitogen-activated protein kiase(MAPK) signaling pathway, phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt) signaling pathway, were mainly associated with radioresistance. Conclusion Multiple genes and signaling pathways are involved in radioresistance, further studies are needed to investigate the radioresistance factors, which could provide new targets for radiotherapy sensitization drugs development.
ABSTRACT
Objective To investigate the antitumor effect of PRDM5 gene in prostate cancer cells. Methods PRDM5 gene was cloned and inserted into lentiviral vector using polymerase chain reaction (PCR), restriction endonuclease and T4 DNA ligase connected method. The lentiviral plasmids carrying PRDM5 gene (LV-PRDM5) or control lentivirus (LV-Luc) were co-transfected with lentiviral packaging plasmid mix into 293T cells by liposome method. The viral supernatants were collected and transduced into human prostate cancer cells 22Rv1. The expression of PRDM5 was verified by Western blotting analysis. The cell proliferation and clone formation ability were detected by cell multiplication and cell cloning experiments. The anchorage independent growth rate of prostate cancer cells was assessed by soft agar colony formation assay. Results The lentivirus vector expressing PRDM5 gene was constructed successfully, and the viral supernatants were obtained. The prostate cancer cell line 22Rv1 stably expressing exogenous PRDM5 was screened and verified by Western blotting analysis. Compared with control cells, the prostate cancer cell line 22Rv1 expressing PRDM5 showed a lower growth rate (multiplication time: [52.5±1.4] vs [44.0±1.3] h), clone formation rate ([1 114±98] vs [1 361±123] colonies per dish) and anchorage independent growth rate ([94.6±8.7] vs [154.0±3.5] colonies per cell, P<0.05). Conclusion Overexpression of PRDM5 has inhibitory effect against proliferation, clone formation and anchorage independent growth of prostate cancer cells in vitro.