1.Akt phosphorylation and mechanism of TRAIL-resistance in A549 cells
Lei LIU ; Yanqiu FANG ; Yaling QI ; Xiaodan LU ; Hanfeng WEI ; Yan TAN
Chinese Journal of Immunology 2014;(5):630-632,638
Objective:To investigate mechanism of TRAIL-resistance in A549 cells ( a cell line of non-small cell lung carcino-ma cells) due to Akt phosphorylation .Methods:A549 cells were treated with Akt inhibitor Perifosine and rsTRAIL protein individual-ly and in combination.The expressions of Akt phosphorylation(p-Akt),c-FLIPLL and caspase-8 were detected by Western blot.The apoptotic rate of the A549 cells treated was detected by flow cytometry and the cell proliferation was evaluated by MTT assay .Results:A549 cells showed the increased level of Akt phosphorylation mediated by rsTRAIL protein .Treatment with the Akt inhibitor Perifosine induced a suppression of Akt activation in A 549 cells and a concomitant decrease in the expression of c-FLIPLL .As a result, Perifosine significantly enhanced TRAIL-induced apoptosis rate of (76.5 ±3.02)%and cytotoxic rate of (83.2 ±2.54)%by promoting the activ-ity of caspase-8.Conclusion:Akt activity promotes A549cells survival against TRAIL-induced apoptosis and that the cytotoxic effect of rsTRAIL protein can be enhanced by modulating the Akt phosphorylation in human non -small cell lung carcinoma cells .
2.Clinical and radiographic evaluation of oblique lumbar interbody fusion combined with percutaneous pedicle screw fixation on computer navigation for lumbar spondylolisthesis
Zhong FANG ; Fang GAO ; Feng LI ; Guangwu LIU ; Jianfeng GUO ; Yong LI ; Yong XU ; Wei WU ; Hanfeng GUAN ; Hui LIAO ; Heng ZENG ; Wei XIONG
Chinese Journal of Orthopaedics 2017;37(16):980-988
Objective To assess the clinical effect of oblique lumbar interbody fusion (OLIF) combined with percutaneous pedicle screw fixation on computer navigation for lumbar spondylolisthesis.Methods Total 20 patients (8 males and 12 females with average age of 54.1± 12.3 years) with lumbar spondylolisthesis were enrolled in our study during Oct.2014 and May.2016.All patients were treated with OLIF combined with percutaneous pedicle screw fixation on computer navigation.Operation time,blood loss and complications were all recorded.Clinical and Radiographic evaluation were investigated on 1 week,3 months,6 months,12 months postoperatively and final follow-up.Visual analogue scale (VAS) for low back pain and leg pain,Oswestry disability index (ODI) for low back pain and the MOS item short form health survey (SF-36) were used to evaluate the clinical efficacy of surgery.Disc height,disc angle,lumbar lordosis and degree of upper vertebral slip of patients were investigated with X-ray.Cross-sectional area of intervertebral foramina was measured with three-dimensional CT and MRI.The cross-sectional area and sagittal diameter of the thecal sac were measured on T2-weighted axial and sagittal magnetic resonance images.Accuracy of pedicle screw placement was investigated with three-dimensional CT.Fusion rate was investigated with three-dimensional CT and Xray.Results All patients were followed for 12-30 months (22.9±4.8 months).The mean operation time was (119.0±23.8) min,the mean blood loss was (57.8±20.6) ml.VAS for low back pain,VAS for leg pain,and ODI were significantly improved from (6.7± 2.6),(6.3±2.7) and 50.5%±18.2% preoperatively to (1.3±1.0),(0.8±1.0) and 14.0%±9.6% at the latest follow-up.The SF-36 PCS and MCS scores were improved from (27.1 ± 13.9) and (51.0±22.7) preoperatively to (67.3± 18.9) and (81.2±14.1) at the latest follow-up.Disc height,disc angle,lumbar lordosis were significantly increased from (6.0±3.6) mm,1.8°±6.2° and 39.2°±8.4° preoperatively to (10.8± 1.7) mm,6.2°±3.5° and 45.0°±7.8° at the latest follow-up.Degree of upper vertebral slip of patients was reduced from 23.5%±7.4% preoperatively to 4.2%±3.1% at the latest follow-up.Cross-sectional area of intervertebral foramina in CT and MRI were significantly increased from (140.6±36.0) mm2 and (78.1±31.2) mm2 before surgery to (179.8±35.6) mm2 and (141.7±29.5) mm2 at 6 months after surgery.Cross-sectional area and sagittal diameter of thecal sac were significantly increased from (73.4±29.3) mm2 and (5.2±3.2) mm before surgery to (124.5±26.6) mm2 and (9.5±2.0) mm at 6 months after surgery.Accuracy of pedicle screw placement was 95%,and fusion rate was 100% at 6 months after surgery.There were no severe vascular and nerve injuries.Conclusion OLIF combined with percutaneous pedicle screw fixation on computer navigation has good indirect decompression effect on lumbar spondylolisthesis,and was associated with high fusion rate.It can also effectively decrease the surgical trauma,improve the accuracy of pedicle screw placement,and increase disc height,disc angle and lumbar lordosis.
3.Expression of circular RNA 102958 in human gastric cancer tissues and its clinical significance
Juan WEI ; Wei WEI ; Qijia ZHANG ; Hanfeng XU
Cancer Research and Clinic 2020;32(6):391-393
Objective:To investigate the expression of circular RNA 102958 (circRNA_102958) in human gastric cancer tissues and its clinical significance.Methods:Thirty cancer tissues from gastric cancer patients in Jiangsu Cancer Hospital from September 2017 to March 2018 and their matched normal gastric mucosa tissue samples were collected. The relative expression of circRNA_102958 in the tissues was detected by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR). The relationship between the expression of circRNA_102958 in gastric cancer tissues and clinicopathological features was analyzed. The receiver operating characteristic curve (ROC) was used to evaluate the efficacy of circRNA_102958 in the diagnosis of gastric cancer.Results:The relative expression of circRNA_102958 in gastric cancer tissues was higher than that in normal gastric mucosa tissues, and the difference was statistically significant (5.1±1.0 vs. 1.0±0.0, t = 4.045, P = 0.000 2). The relative expression of circRNA_102958 in gastric cancer tissues of patients with stage Ⅲ-Ⅳ was higher than that of patients with stage Ⅰ-Ⅱ, and the difference was statistically significant (9.3±2.6 vs. 2.0±0.5, t = 2.302, P = 0.029). There was no statistical difference in the relative expression of circRNA_102958 among patients with different gender, age, the longest tumor diameter, tissue differentiation degree, and lymph node metastasis (all P > 0.05). ROC analysis showed that the sensitivity and specificity of circRNA_102958 in the diagnosis of gastric cancer were 74% and 61%, and the area under the curve was 0.74. Conclusions:circRNA_102958 is highly expressed in gastric cancer tissues, and its expression is related to the stage of gastric cancer, which may be related to the occurrence and development of gastric cancer. circRNA_102958 is expected to become a molecular marker for gastric cancer diagnosis.
4.The characteristics of hypoxia-induced exosomes secreted from A549 cells and its effect on radioresistance
Chuandong ZHU ; Lixue WANG ; Guoxiang WANG ; Juan WEI ; Qijia ZHANG ; Hanfeng XU ; Jinlong TONG ; Qin ZHENG
Chinese Journal of Radiological Medicine and Protection 2018;38(8):567-573
Objective To investigate the effect of exosomes secreted from human lung adenocarcinoma A549 cells under hypoxic or normoxic conditions on the radiosensitivity and invasiveness of normoxia cells.Methods A549 cells were cultured in hypoxic (1% O2) and normoxic (21% O2) conditions,respectively.The exosomes (N-EXO and H-EXO) secreted from normoxic or hypoxic A549 cells were collected by ultracentrifugation and its number was measured using a NanoSight detector.The appearance and size distribution of exosomes were observed by a scanning electron microscopy.The exosomal marker protein CD63 was measured by Western blot.The proliferation of cells exposed to X-rays under hypoxic or normoxic conditions were detected by CCK8 assay.The cell uptake situation of exosomes labeled with PKH67 was observed by a fluorescence microscopy.Cell migration and invasiveness were detected by a cell scratch test and transwell assay.The expression of matrix metalloproteinase 2 (MMP2) and MMP9 was detected by ELISA.Cellular radioresistance effect of exosomes was evaluated by a colony formation assay.Results The NanoSight measurement showed the number of exosomes in cell culture medium was increased after hypoxia treatment.The H-EXO and N-EXO showed typical ring cake shape.The size distribution of H-EXO was mainly between 30 nm and 200 nm,smaller than that of N-EXO (50-220 nm).Western blot assay showed that CD63 was expressed in both H-EXO and N-EXO.At 4 and 6 days after 2 Gy X-rays irradiation,cell proliferation rate of hypoxia A549 cells was significantly higher than that of normoxia cells.The green fluorescent marker of exosomes,PKH67,was distributed inside of the cell.Cell scratch test showed that the width of H-EXO group was much smaller than that of N-EXO group at 12,24 and 48 hours after exosomes treatment (t =2.96,6.76,3.35,P < 0.05).Transwell assay showed that the number of transmembrane cells in the H-EXO group was more than that in the N-EXO group and the control group (t =4.84,7.88,P < 0.O1).The expression levels of MMP2 (t =4.70,3.21,P<0.05) and MMP9 (t =5.61,3.76,P<0.05) in the supernatant of H-EXO group were significantly higher than those in the control and N-EXO groups.Cell survival assay showed that the D0 values of control,N-EXO and H-EXO group were 2.614,2.552 and 4.50 respectively,indicating that H-EXO could enhance radioresistance of A549 cells significantly.Conclusions This study finds that the number of exosomes released from A549 cells was increased under hypoxic condition but its size becomes smaller than that under normoxia.Hypoxic exosomes can promote the migration of normoxia cells andenhance cell radioresistance as well.