1.Effects of CENP-A on invasion and migration of ovarian cancer cells by regulating PI3K/AKT/NF-κB signaling pathway
Guohua CHENG ; Yarong LI ; Zhaoyu LIU ; Xiang SONG
Chinese Journal of Endocrine Surgery 2021;15(3):248-253
Objective:To investigate the effects of centromere protein-A (CENP-A) on the invasion and migration of ovarian cancer (OC) cells and explore the related mechanism.Methods:OC cell line A2780 was cultured in vitro, and they were divided into Ng Group (Blank Control Group) , pcDNA group (negative transfection group:PCDNA vector plasmid) , pcDNA-CENP-A group (over-expression Group: pcDNA-CENP-A Vector Plasmid) and pathway inhibitor group (TRANSFECTION-CENP-A+ PI3K pathway inhibitor LY294002) . The cell proliferation was detected by CCK-8 method; the cell migration and invasion was detected by Scratch test and Transwell test; the expression of CENP-A, E-cadherin, N-cadherin and phosphatidylinositol 3-kinase/protein kinase B/nuclear factor-kappa B (PI3K/AKT/NF-κB) pathway related proteins was detected by Western blot.Results:A2780 cells were successfully transfected. After 24 hours, with the extension of culture time, compared with that in NG group [ (0.50±0.07) , (0.72±0.11) , (0.99±0.14) ] and pcDNA group [ (0.55±0.08) , (0.78±0.12) , (1.02±0.15) ], the viability of A2780 cells in pcDNA-CENP-A group [ (0.78±0.12) , (1.03±0.15) , (1.67±0.25) ] and pathway inhibitor group [ (0.63±0.09) , (0.87±0.13) , (1.39±0.20) ] increased significantly ( P<0.05) , compared with that in the pcDNA-CENP-A group, the viability of A2780 cells in the pathway inhibitor group was significantly decreased ( P<0.05) , in a time-dependent manner. Compared with those in NG group [ (15.83±1.46) %, (105.32±15.78) individual] and pcDNA group [ (16.79±1.46) %, (108.98±16.35) individual], the migration rate [ (37.96±5.80) %, (25.15± 2.19) %] and invasion number [ (327.87±49.18) individual, 206.53±30.97) individual] of A2780 cells, protein expression of CENP-A, N-cadherin, Vimentin, p-PI3K/PI3K, p-AKT/AKT, NF-κB, interleukin (IL-1β) , tumor necrosis factor-α (TNF-α) in pcDNA-CENP-A group and pathway inhibitor group were significantly higher ( P<0.05) , the expression of E-cadherin was significantly lower ( P<0.05) ; compared with those in the pcDNA-CENP-A group, the migration rate and invasion number of A2780 cells, protein expression of CENP-A, N-cadherin, Vimentin, p-PI3K/PI3K, p-AKT/AKT, NF-κB, interleukin (IL-1β) , tumor necrosis factor-α (TNF-α) in pathway inhibitor group were significantly lower ( P<0.05) , and the expression of E-cadherin was significantly higher ( P<0.05) . Conclusion:Overexpression of CENP-A can promote the proliferation, invasion and migration of ovarian cancer cells, which may be achieved by activating PI3K/AKT/NF-κB signaling pathway.
2.Analysis of lens power and its influencing factors in children aged 3-12 years
Zhaoyu XIANG ; Qiurong LIN ; Yan XU ; Jun QIANG ; Yu QIAN ; Haidong ZOU
Chinese Journal of Experimental Ophthalmology 2021;39(6):550-556
Objective:To understand the changing trend and influencing factors of lens power (LP) in children aged 3-12 in Jing'an District, Shanghai.Methods:A cross-sectional study was conducted.One hundred and thirty-one eyes of 131 patients with refractive errors were included in the Optometry Clinic of Shanghai Eye Hospital from October 2019 to January 2020.The 1% atropine sulfate was employed to dilate pupils for children aged 6 years or younger, and 0.5% topiramate for children older than 6 years.The axial length, mean keratometry (Km), anterior chamber depth (ACD), lens thickness (LT) and central corneal thickness (CCT) were measured using an IOL Master.The spherical equivalent (SE) and best corrected visual acuity (BCVA) were measured after cycloplegia using autorefractor and phoropter, and the LP was calculated using the Bennett formula.The patients were divided into different age groups, including 3-4 years group (16 eyes), 5-6 years group (20 eyes), 7-8 years group (25 eyes), 9-10 years group (33 eyes) and 11-12 years group (37 eyes). There were 57 eyes in the male group and 74 eyes in the female group.The patients were also divided into different refractive groups, including mild myopia group (38 eyes), moderate myopia group (12 eyes), high myopia group (25 eyes), emmetropia group (11 eyes), mild hyperopia group (9 eyes), moderate hyperopia group (13 eyes), and high hyperopia group (23 eyes). The differences in ocular biological parameter measurements between different age groups, different gender groups and different refractive groups were compared and the correlations between age, eye parameters and LP were analyzed using Pearson correlation analysis.The contribution of multiple influencing factors to LP was analyzed by multiple linear regression models.The study protocol followed the Declaration of Helsinki and was approved by an Ethics Committee of Shanghai General Hospital, Shanghai Jiao Tong University Hospital (No.2020KY018). Written informed consent was obtained from each guardian of the subject.Results:The average LP of children in the 3-4 years group, 5-6 years group, 7-8 years group, 9-10 years group and 11-12 years group were (27.35±1.88), (24.71±1.92), (22.92±1.87), (21.49±1.54) and (21.25±1.55) D, respectively.With the increase of age, the LP value was decreased gradually.There were significant differences between 3-4 years group and 5-6 years group, 5-6 years group and 7-8 years group, 7-8 years group and 9-10 years group (all at P<0.05). The average LP value of girls was obviously higher than that of boys ( t=-3.38, P<0.01). The LP value of the high myopia group was significantly lower than that of the emmetropia group, and the LP values of the moderate myopia and the low myopia group were significantly lower than that of the hyperopia group, and the LP values of the low hyperopia group and the moderate hyperopia group were significantly higher than that of the emmetropia group (all at P<0.05). The LP value was negatively correlated with age, AL, ACD and CCT ( r=-0.76, -0.79, -0.38, -0.18; all at P<0.05), and was positively correlated with SE and LT ( r=0.62, P<0.05; r=0.68, P<0.01). There was no obvious correlation between Km and LP ( r=0.07, P=0.45). The independent influencing factors of LP were analyzed through multiple linear regression equations, showing that LP=-0.430×AL+ 0.329×LT-0.267×age-0.108×gender-0.084×CCT (male=1, female=0). The standardized coefficients of each factor arranged in descending order were AL, LT, age, gender and CCT (all at P<0.05). Conclusions:The LP of children aged 3-12 in Jing'an District of Shanghai decreases with age and increases with SE.LP values of girls are higher than those of boys.