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Objective:To observe the effect of Gemcitabine ( GEM) on the viability and apoptosis of non-small cell lung cancer HCC827 in vitro. Methods:The cell viability,apoptosis and cell cycle of HCC827 cells induced by Gemcitabine were detected with cell counting kit-8 assay (CCK-8),Annexin V-FITC/PI staining and flow cytometry. The expression of Bcl-2 protein of cells treated with GEM was examined by Western blot assay. Results: There was significant inhibition effect on HCC827 cells treated with 0. 1-1 000 ng/ml of GEM,which can promote the occurrence of HCC827 cell apoptosis and arrest cell in the S phrase. The apoptosis induced by GEM was accompanied with the down regulation of Bcl-2 protein. Conclusion: GEM can inhibit the cell viability and induce the HCC827 cell apoptosis and S phrase arrest. Its cell dead type was apoptosis,which was related with the expression of Bcl-2 protein.
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[ ABSTRACT] AIM:To investigate the effect of HOX transcript antisense RNA ( HOTAIR) on the migration and invasion abilities of liver carcinoma HepG 2 cells.METHODS:The expression of phosphoinositide-3-kinase regulatory sub-unit 3 (PIK3R3) in the liver cancer and normal liver tissues was detected by immunohistochemistry .The efficiency of gene silencing of HOTAIR or PIK3R3 by LV3-shHOTAIR or LV3-shPIK3R3 was determined by qPCR and Western blot .The mi-gration and invasion abilities of HepG 2 cells after silencing of HOTAIR and PIK3R3 were measured by wound healing assay and Transwell Matrigel invasion assay .The expression of miR-214 after silencing of HOTAIR and PIK3R3 was analyzed by qPCR.The expression of HOTAIR and PIK3R3 in the HepG2 cells was also evaluated by qPCR after transfected with miR-214 mimics or miR-214 inhibitor .Dual-luciferase reporter assay system was used to determine the regulatory effect of miR-214 on HOTAIR and PIK3R3 expression.RESULTS:PIK3R3 expression increased significantly in the liver cancer tissues compared with normal liver tissues .The abilities of invasion and metastasis of hepatocellular carcinoma were reduced after silencing of HOTAIR and PIK3R3.miR-214 expression was increased when silencing of HOTAIR and PIK3R3 was per-formed.HOTAIR and PIK3R3 expression was reduced after transfection with miR-214 mimics.HOTAIR and PIK3R3 ex-pression was increased after transfection with miR-214 inhibitor.The results of dual-luciferase reporter assay test showed that miR-214 directly regulated HOTAIR and PIK3R3 transcription activity .CONCLUSION: HOTAIR regulates the ex-pression of PIK3R3 through miR-214, thus promoting the migration and invasion abilities in the liver cancer cells .
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Objective To investigate the effects of mannan-binding lectin (MBL) on IL-8 and TNF-α production induced by Candida albicans ( C. albicans) in human THP1/CD14 monocytes. Methods The THP1/CD14 cells were stimulated for 24 h with heat-inactivated yeast form or hyphal form cells of C. albicans strain at the indicated ratios after pretreated with human natural MBL at concentrations ranging from 1 to 20 mg/L for 2 h. The content of IL-8 and TNF-α in culture supernatants were detected by ELISA,and the levels of IL-8 and TNF-α mRNA expressions in these cells were determined by RT-PCR. Western blot was used to detect C. albicans-induced NF-κB translocation in THP1/CDI4 cells. Results ELISA showed that secretion of IL-8 and TNF-α from THP1/CD14 cells could be induced by both yeast cells and hyphal cells. Hyphal cells proved to be much less efficient than yeast cells in stimulating production of IL-8and TNF-α by THP1/CD14 cells. The productions of IL-8 and TNF-α by THP1/CD14 cells induced with C.albicans were profoundly inhibited by MBL at higher concentrations ( 10-20 mg/L) but not MBL at lower concentrations ( 1 mg/L). RT-PCR analysis also indicated that the mRNA expressions of IL-8 and TNF-αt in THP1/CD14 cells were decreased to various extents by MBL at higher concentration, compared to the corresponding THP1/CD14 cells stimulated with C. albicans only. Similarly, MBL at higher concentration ( 20mg/L) decreased the NF-κB translocation in THP1/CD14 cells. Conclusion MBL may inhibit IL-8 and TNF-α production induced by dimorphism C. albicans in THP1/CD14 cells, suggesting that MBL can play some roles on the regulation of C. albicans immune response.