1.Effect of Toll like-receptor 4 on proliferation of cervical cancer cells by NF-κB signaling pathway
Wei YAN ; Fanggen XU ; Anwen LIU ; Jing CAI ; Shuo WANG
Chinese Journal of Pathophysiology 2015;(2):301-307
AIM:To investigate whether specific small interfering RNA ( siRNA) targeting Toll-like receptor 4 ( TLR4) gene inhibits the proliferation of cervical cancer cell line HeLa through NF-κB signaling pathway and down-regula-tion of Bcl-2 expression.METHODS:Three specific TLR4 siRNAs and 1 negative siRNA were transfected respectively in-to HeLa cells.The expression of TLR4 at mRNA and protein levels was measured by reverse transcription-polymerase chain reaction ( RT-PCR) and Western blotting , respectively .The protein expression of p 65 and Bcl-2 was detected by Western blotting.The cell proliferation was determined by MTT assay and plate colony formation assay .The apoptotic rate of the cells and the cell cycle were analyzed by flow cytometry .RESULTS:In comparison with the other 2 kinds of TLR4 siR-NAs, TLR4-siRNA-003 demonstrated the strongest silencing effect on TLR4 gene expression in the HeLa cells at 48 h after transfection.The expression of TLR4 at mRNA and protein levels was reduced by 62% and 89%, respectively.The pro-tein levels of p65 and Bcl-2 significantly decreased .The growth rate of HeLa cells transfected with TLR 4-siRNA-003 was significantly inhibited .Moreover , the cell apoptotic rate increased significantly and the cell cycle was arrested at G 1 phase as the HeLa cells were transfected with TLR 4-siRNA-003 for 48 h.CONCLUSION:Specific TLR4 siRNA effectively in-hibits the expression of TLR4 and inhibits the proliferation of cervical cancer HeLa cells through NF-κB signaling pathway and down-regulation of Bcl-2 expression, indicating that TLR4 may be a new target for the treatment of cervical cancer .
2.Mobilization of Peripheral Blood Stem Cells with High Dose Cyclophosphamide Combination Chemotherapy and G-CSF in Breast Cancer Patients
Shikai WU ; Santai SONG ; Xiaoqing LIU ; Zefei JIANG ; Anwen YAN ; Wenhu WANG ; Jingxin YU ; Yimei QU
Journal of Experimental Hematology 2000;8(4):295-298
To evaluate the effect of mobilization of peripheral blood stem cells (PBSC) with high dose cyclophosphamide combination chemotherapy and G-CSF in breast cancer patients, a new mobilization protocol was designed on the basis of standard combination chemotherapy regimen, in which the dose of cyclophosphamide was raised to 2 to 4 times, and G-CSF began to be used at the dose of 150 micro g twice everyday when white blood cell (WBC) decreased below 1.0 x 10(9)/L. PBSC collection was performed while WBC increased over 5.0 x 10(9)/L during bone marrow recovering. The PBSC mobilization protocol was completed in 10 patients, the median nadir of WBC was 0.8 (0.4 - 1.0) x 10(9)/L, the median time of PBSC collection was 2 (2 - 4), the median number of collected CD34(+) cells was 6.43 (1.99 - 8.75) x 10(6)/kg. The results showed that the protocol, high dose cyclophosphamide combination chemotherapy, was an optimal PBSC mobilization regimen in breast cancer patients.