1.Effect of Rab11 gene expression on the invasion and migration of cervical cancer cell line SiHa in hypoxia
Hao XU ; Yuan YUAN ; Jiayin JI ; Qian JIANG ; Linjun NIU ; Nianli LIU ; Longzhen ZHANG ; Xia WANG
Chinese Journal of Obstetrics and Gynecology 2016;51(12):928-933
Objective To explore the expression of Ras-related protein 11(Rab11)in hypoxia, the effect of Rab11 on the invasion and migration of cervical cancer cell line SiHa and its possible mechanism. Methods SiHa cells were divided into 4 groups, the normoxic blank group (normal culture in normoxia), the hypoxic blank group (normal culture in hypoxia), the negative control group [transfection of negative control small interfering RNA(siRNA)in hypoxia], the Rab11-siRNA group (transfection of Rab11 siRNA in hypoxia). Western blot was used to examine the expression of Rab11, integrin α5, integrin β3, phosphorylated focal adhesion kinase(p-FAK), phosphorylated phosphatidylinositol 3 kinase(p-PI3K) protein, together with the expression of Ras correlative C3 creotoxin substrate 1(Rac1), which was critical in regulating cell invasion. The mRNA expression of Rab11 in the 4 groups was detected by realtime-qPCR. The cell invasion was detected by matrigel assay, while the cell migration was detected by transwell assay. Immunofluorescence was used to identify intracellular location of Rac1 in SiHa cell. Results (1) The expression of Rab11, intergrin α5, intergrin β3, p-FAK, p-PI3K and Rac1 in the normoxic blank group were 0.56±0.04, 0.33±0.03, 0.32±0.03, 0.36±0.03, 0.35±0.03 and 0.47±0.03, respectively. In the hypoxic blank group, they were 0.73±0.03, 0.74±0.03, 0.61±0.03, 0.62±0.03, 0.60±0.03 and 0.73±0.03, respectively. In the negative control group, their expressions were 0.72±0.03, 0.73±0.03, 0.59±0.03, 0.61±0.03, 0.59±0.03 and 0.72±0.03, respectively. While in the Rab11-siRNA group, they were 0.44±0.03, 0.30±0.03, 0.29±0.03, 0.30±0.03, 0.30±0.03 and 0.34±0.04, respectively. The expressions of Rab11, α5, β3, p-FAK, p-PI3K and Rac1 were significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and were significantly lower in the Rab11-siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (2) The expressions of Rab11-mRNA were 1.000±0.000, 1.454±0.114, 1.442±0.101, 0.570± 0.046 in the normoxic blank group, the hypoxic blank group, the negative control group and the Rab11- siRNA group, respectively. It was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11-siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (3) By Matrigel, the invasion cell number in the normoxic blank group, the hypoxic blank group,the negative control group and the Rab11-siRNA group were 65±12, 106±16, 104± 17 and 50±11, respectively. The invasion capacity was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11- siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (4) By transwell assay, the migration cells in the normoxic blank group, the hypoxic blank group, the negative control group and the Rab11-siRNA group were 127±12, 169±15, 161±13 and 77±13, respectively. The capacity of invasion was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11- siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (5) The immunofluorescence showed that the red fluorescence intensity around nucleus was significantly increased in the normoxic blank group, the hypoxic blank group and the negative control group than in the Rab11- siRNA group. Conclusions Hypoxia could promote the invasion and migration of SiHa cells. In hypoxia, the down regulation of Rab11 expression could inhibit the invasion and migration of SiHa cells. This might be due to the decreased expression of the intergrin α5, intergrin β3, p-FAK, p-PI3K and Rac1 protein.
2.Effects of down-regulating peroxiredoxin 2 on the invasion, migration and proliferation of human gastric cancer MGC803 cells
Wenqian WU ; Hao XU ; Renxian TANG ; Kuiyang ZHENG ; Nianli LIU
Cancer Research and Clinic 2017;29(12):793-798
Objective To explore the effects of the down-regulating peroxiredoxin 2 (PRDX2) on invasion, migration and proliferation of human gastric cancer MGC803 cells by using RNA interference. Methods MGC803 cells were divided into 3 groups:blank control group,negative control group and PRDX2 siRNA group. Transwell assay was used to examine the invasive ability change of MGC803 cells after transfection. Scratch test was used to detect the change of MGC803 cells migration ability after transfection. Western blot was used to examine the expression changes of PRDX2, matrix metalloproteinase (MMP)-2 and MMP-9. The proliferation ability of MGC803 cells was assessed by using CCK-8 assay. Results The expressions of PRDX2(0.345±0.006,0.721±0.013,0.720±0.014),MMP-2(0.067±0.012, 0.391±0.015, 0.371± 0.016) and MMP-9 (0.073±0.013, 0.341±0.028, 0.346±0.024) in the PRDX2 siRNA group were lower than those in the blank control group and negative control group (all P < 0.05). The cell invasion, migration and proliferation were inhibited in MGC803 cells (all P < 0.05). Conclusion PRDX2 is overexpressed in MGC803 cells. Down-regulating the expression of PRDX2 could inhibit the invasion, migration and proliferation of MGC803 cells.
3.Effectiveness of primary care quality improvement project in general practice standardized residency training
Nianli ZHOU ; Hui WEN ; Lei JIANG ; Yun LIU ; Meng ZHANG ; Wen PENG
Chinese Journal of General Practitioners 2024;23(1):61-64
A 6-month primary care quality improvement (QI) project was conducted for 63 general practice residents at Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from November 2021 to April 2022. The effectiveness of the QI project on the post competency of general practice residents was comprehensively assessed by three dimensions: self-satisfaction, objective evaluation and teacher-evaluation. The overall satisfaction score of general practice residents was significantly increased after the implementation of QI project((3.83±0.67) vs. (3.41±0.63), t=3.35, P=0.009). The total score of objective assessment was increased from (73.48±8.04) before the project implementation to (78.14±5.24) after the implementation ( t=3.37, P=0.001). The total score of training effectiveness significantly increased from (57.57±11.84) before the project implementation to (79.27±8.40) after the implementation ( t=30.07, P<0.001). The results indicate that the primary care QI project can improve the post competency of general practice residents, and also improve the self-satisfaction of residents for active learning and participation in the training.