1.Expression and regulatory mechanism of S100A14 in breast cancer
Chengshan XU ; Hongyan CHEN ; Chengrong LU ; Zhihua LIU
Chinese Journal of Oncology 2016;38(4):252-257
Objective To explore the expression of S100A14 in breast cancer tissue, and the EGF and S100A14 feedback regulatory mechanism. Methods S100A14 mRNA level in 52 cases of of breast cancer and adjacent normal tissue was detected by quantitative real?time PCR. S100A14 protein in 21 cases of breast cancer and adjacent normal tissue was detected by Western blot. S100A14 mRNA after EGF treatment was detected by RT?PCR and real?time PCR. The levels of S100A14, p?ERK and t?ERK were detected by Western blot. Knocking down S100A14 expression was performed by siRNA technology. Results The levels of S100A14 mRNA and protein were significantly increased in breast cancer tissues ( P<0.05 for both) . The high expression of S100A14 was related with the recurrence of breast cancer patients ( P=0.038). S100A14 mRNA level was significantly up?regulated in the MDA?MB?453 cells (1.50±0.11) and MCF?7 cells (1.40±0.03) after 1 ng/mL EGF treatment, and 1.66±0.08 and 1.71±0.17 in the MDA?MB?453 cells after 10 ng/mL EGF treatment, significantly higher than that of the control group (1.00±0.09 and 1.00±0.03) (P<0.05 for both). In the TD47 cells, the S100A14 mRNA levels in the control, 1 ng/ml EGF and 10 ng/ml EGF + U0126 treatment groups were 1. 00 ± 0. 04, 1. 56 ± 0. 04 and 1. 00 ± 0. 10, respectively ( P<0.05) . Conclusions The expression of S100A14 mRNA and protein is promoted by EGF through p? ERK signaling pathway in breast cancer cells. There may be a feedback loop between EGF and S100A14.
2.Expression and regulatory mechanism of S100A14 in breast cancer
Chengshan XU ; Hongyan CHEN ; Chengrong LU ; Zhihua LIU
Chinese Journal of Oncology 2016;38(4):252-257
Objective To explore the expression of S100A14 in breast cancer tissue, and the EGF and S100A14 feedback regulatory mechanism. Methods S100A14 mRNA level in 52 cases of of breast cancer and adjacent normal tissue was detected by quantitative real?time PCR. S100A14 protein in 21 cases of breast cancer and adjacent normal tissue was detected by Western blot. S100A14 mRNA after EGF treatment was detected by RT?PCR and real?time PCR. The levels of S100A14, p?ERK and t?ERK were detected by Western blot. Knocking down S100A14 expression was performed by siRNA technology. Results The levels of S100A14 mRNA and protein were significantly increased in breast cancer tissues ( P<0.05 for both) . The high expression of S100A14 was related with the recurrence of breast cancer patients ( P=0.038). S100A14 mRNA level was significantly up?regulated in the MDA?MB?453 cells (1.50±0.11) and MCF?7 cells (1.40±0.03) after 1 ng/mL EGF treatment, and 1.66±0.08 and 1.71±0.17 in the MDA?MB?453 cells after 10 ng/mL EGF treatment, significantly higher than that of the control group (1.00±0.09 and 1.00±0.03) (P<0.05 for both). In the TD47 cells, the S100A14 mRNA levels in the control, 1 ng/ml EGF and 10 ng/ml EGF + U0126 treatment groups were 1. 00 ± 0. 04, 1. 56 ± 0. 04 and 1. 00 ± 0. 10, respectively ( P<0.05) . Conclusions The expression of S100A14 mRNA and protein is promoted by EGF through p? ERK signaling pathway in breast cancer cells. There may be a feedback loop between EGF and S100A14.
3.K562 cells induces apoptosis of activated NK cells in vitro.
Yan CAO ; Lianning DUAN ; Chengrong LU ; Yuan LUO ; Peide XIANG ; Shu YAN ; Shujing GE ; Yanjun ZHANG
Journal of Southern Medical University 2012;32(9):1245-1249
OBJECTIVETo investigate the apoptosis of NK cells induced by the erythroleukemia cell line K562 in vitro.
METHODSPrimary NK cells isolated from the peripheral blood of healthy donors by magnetic-activated cell sorting were cultured with stem cell medium containing recombinant human interleukin-2 (rhIL-2). The NK cells and K562 cells were mixed and co-cultured at different E:T ratios for different time lengths. The apoptosis of NK cells and K562 cells were detected using PE-AnnexinV/7-AAD labeling and flow cytometry.
RESULTSThe purity of isolated NK cells reached (93.99∓4.22)%. At the same E: T ratio, the apoptotic rate of NK cells induced by K562 cells increased significantly with time. As the E:T ratio reduced, the apoptotic rate of the NK cells increased and their cytotoxic activity against K562 cells was attenuated.
CONCLUSIONK562 cells can induce the apoptosis of activated NK cells, which is one of the probable mechanisms of immune escape of tumors.
Apoptosis ; Cytotoxicity, Immunologic ; Humans ; K562 Cells ; Killer Cells, Natural ; cytology ; immunology ; Tumor Escape
4.Screening of pregnane X receptor activation from ginsenosides.
Yuguang WANG ; Haosheng LIU ; Xianxie ZHANG ; Yong XIAO ; Beibei LU ; Zengchun MA ; Qiande LIANG ; Xianglin TANG ; Chengrong XIAO ; Hongling TAN ; Boli ZHANG ; Yue GAO
Acta Pharmaceutica Sinica 2013;48(1):144-8
In order to study effects of ginseng on the metabolism of drug belong to CYP3A4 substrate, screening of pregnane X receptor activation from ginsenosides was performed by reporter assay. Based on PXR-CYP3A stable translation cell lines, 13 ginsenosides were screened for pregnane X receptor activation by reporter assays, and RIF as the positive control. The effect of ginsenosides Rg1 onCYP3A4 mRNA expression was also investigated by RT-PCR. The PXR-CYP3A stable translation cell lines had good response to RIF, and the EC50 is 2.51 micro mol x L(-1). When the condition of final concentration was 10 micromol x L(-1), ginsenoside F2 and protopanaxatriol had moderate inductive effects on PXR. Panaxotriol, Rg2, pseudoginsenoside F11, Rg1, ginsenoside and Rb3 had inhibitory effects on PXR. Ginsenoside Rf1, Rg3, Rh2 and protopanaxdiol had no obvious effects on PXR. Rg1 down-regulated CYP3A4 mRNA expression in a concentration-dependent manner. Activation of pregnane X receptor by ginsenosides may influence the metabolism of drug belong to CYP3A4 substrate, and cause ginseng-drug interactions.
5.Clinical study of persistent abnormal muscle response after microvascular decompression for hemifacial spasm
Chengrong JIANG ; Wu XU ; Tianyu LU ; Yuxiang DAI ; Chen YU ; Hongbin NI ; Weibang LIANG
Chinese Journal of Neuromedicine 2017;16(10):994-998
Objective To explore the causes of persistent abnormal muscle response (AMR) after microvascular decompression (MVD) for hemifacial spasm (HFS) and the clinical outcomes of these patients.Methods MVD was performed under intraoperative electrophysiological monitoring of AMR in 372 HFS patients in 2014.Before MVD,the characteristic AMR of HFS was recorded in 326 patients.The patients were divided into two groups based on whether AMR disappeared or persisted following MVD;21 patients showed persistent AMR after successful MVD while AMR disappeared after decompression in the other 305 patients.The clinical features,treatment efficacy and postoperative complications were compared between these two groups.Results Gender,side of depression and mean age between the two groups showed no significant differences (P>0.05).The immediate postoperative cure rate of the AMR disappeared group (88.9%) was significantly higher than that in the AMR persisted group (28.6%,P<0.05).The follow-up cure rate showed no significant difference between the two groups (P>0.05),and the postoperative and follow-up complications showed no significant differences (P>0.05).Conclusion The long duration of HFS patients may be responsible for persistent AMR after successful decompression,and it is more likely for these patients to get delayed cured;their long-term outcomes showed no difference as compared with those in patients with disappeared AMR after MVD.