1.Study on effect mechanism of sanguinarine on proliferation, apoptosis, invasion and migration of cervical cancer cells
Zhencai LI ; Ping JIANG ; Qiuyu WANG ; Li YANG ; Pengxiao FU ; Jinru ZHANG
Chongqing Medicine 2017;46(22):3039-3042
Objective To investigate the effect mechanism of sanguinarine on the proliferation,apoptosis,invasion and migration abilities of cervical cancer cells.Methods MTT,flow cytometer,cell scratch test and Transwell chamber assay were respectively used to detect the cellular proliferation,apoptosis,migration and invasion abilities after sanguinarine action.The expression levels of E-Cadherin,PTEN,β-catenin and MMP2 protein of cervical cancer cells after sanguinarine action were detected by Western blot.Results 0.6,0.8 μmol/L sanguinarine had the inhibitory effect on the proliferation of cervical cancer cells.After 0.8μmol/L sanguinarine action for 48 h,cervical cancer HeLa and Siha cells apoptosis rate were up to (45.68± 2.26)% and(31.89 ± 3.80)% respectively.0.8 μmol/L sanguinarine action for 3 h,cervical cancer cells HeLa and Siha adhesion rates were only (67.45 ± 2.13)%and(73.59± 2.61)%.0.8 mol/L sanguinarine action for 16 h,the invasion numbers of cervical cancer Hela and Siha cell were (39.64 ±1.98) and (43.87 ± 2.83) respectively.The expression amount of E-Cadherin and PTEN in cervical cancer cells after sanguinarine action was increased,while the expression amount of E-Cadherin and PTEN was weakened.Conclusion Sanguinarine has the proliferation inhibiting and apoptosis promoting effect on cervical cancer cells,its mechanism may be related to adhesion protein E-Cadherin,β-catenin and PTEN,MMP2.
2.miR-139-5p inhibits proliferation and invasion of epithelial ovarian cancer cells by targeting Notch1
JIANG Ping ; YANG Xike ; WANG Qiuyu ; SHAO Lanyun ; WANG Songpeng ; FANG Jianrui ; FU Pengxiao ; GUO Yingying
Chinese Journal of Cancer Biotherapy 2020;27(1):19-24
Objective: To explore the action mechanism of miR-139-5p inhibiting proliferation and invasion of epithelial ovarian cancer (EOC) cells by targetedly regulatingNotch1.Methods: A total of 24 pairs of EOC tissues and its corresponding para-cancerous tissues from patients, who underwent surgical resection in the DepartmentofGynecology,Nanyang Central Hospital of Henan Province, were collected for this study; in addition, human ovarian cancer cell lines (SKOV3, ES2, HEY-T30) and human ovarian epithelial cell line IOSE80 were also collected. Real-time quantitative PCR (qPCR) was applied to detectmRNAexpressionofmiR-139-5pandNotch1 in EOC tissues and cell lines. The miR-139-5p over-expression vector and recombinant plasmid pLV-Notch1 were transfected into SKOV3 cells. Blank control group (Ctrl group) and negative control group (NC group) were set up. Dual luciferase reporter gene assay was applied to verify the targeting relationship between miR-139-5p and Notch1 3'-UTR. CCK-8, Transwell and Scratch healing experiments were applied to detect cell proliferationinvasionandmigration, respectively. Western blotting was applied to detect expressions of proliferation and migration related proteins in cells. Results: Compared with para-cancerous tissues and IOSE80 cells, the expression of miR-139-5p was significantly decreased in EOC tissues and cell lines, while the expression of Notch1 mRNA was significantly increased (all P<0.01). The results of Dual luciferase reporter showed that Notch1 was the downstream target gene of miR-139-5p. Compared with NC group, cell proliferation, invasion and migration ability, expression levels of Notch1, NICD, Cyclin D1, Cyclin A1, Snail1, β-catenin and N-cadherin were all significantly decreased on 3 d in miR-139-5p mimic group (all P<0.01), while expression of E-cadherin was significantly increased (P<0.01); meanwhile, over-expression of Notch1 could reverse the inhibitory effect of miR-1395p on proliferation, invasion and migration of SKOV3 cells. Conclusion: miR-139-5p can targetedly regulate Notch1 to inhibit proliferation, invasion and migration of EOC cells, which may be related to its down-regulation of NICD, Cyclin D1, Cyclin A1, Snail1, βcatenin and N-cadherin, and up-regulation of E-cadherin.
3.Mitochondria-The intracellular receptors for spatial microgravity
Lerong CHEN ; Pengxiao WANG ; Zhen WANG ; Jiangang LONG
Space Medicine & Medical Engineering 2024;35(4):258-267
The weightlessness effect during space flight can cause damage to the astronaut's body,and this damage requires a longer recovery time even after the astronaut returns to Earth,and even irreversible damage may occur.Mitochondria are important organelles within cells,participating in regulating a range of important physiological and biochemical processes.Mitochondrial mediated metabolic regulation can sense microgravity stress,which is an early intracellular response caused by microgravity,promoting the adaptation of intracellular metabolism and stress-induced damage.This article reviews the research progress on the effects and related mechanisms of microgravity and simulated microgravity on cell mitochondria,providing a theoretical basis for the development of protection strategies targeting mitochondria.
4.Modified Xiaoyaosan Alleviates Depression-like Behaviors by Regulating Activation of Hippocampal Microglia Cells in Rat Model of Juvenile Depression
Jiayi SHI ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Feng QIU ; Chang LEI ; Hongyu ZENG ; Kaimei TAN ; Hongqing ZHAO ; Dong YANG ; Yuhong WANG ; Pengxiao GUO ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):46-56
ObjectiveTo investigate the mechanism of Baihuan Xiaoyao Decoction (Xiaoyaosan added with Lilii Bulbus and Albiziae Cortex) in alleviating depression-like behaviors of juvenile rats by regulating the polarization of microglia. MethodSixty juvenile SD rats were randomized into normal control, model, fluoxetine, and low-, medium-, and high-dose (5.36, 10.71, 21.42 g·kg-1, respectively) Baihuan Xiaoyao decoction groups. The rat model of juvenile depression was established by chronic unpredictable mild stress (CUMS). The sucrose preference test (SPT) was carried out to examine the sucrose preference of rats. Forced swimming test (FST) was carried out to measure the immobility time of rats. The open field test (OFT) was conducted to measure the total distance, the central distance, the number of horizontal crossings, and the frequency of rearing. Morris water maze (MWM) was used to measure the escape latency and the number of crossing the platform. The immunofluorescence assay was employed to detect the expression of inducible nitric oxide synthase (iNOS, the polarization marker of M1 microglia) and CD206 (the polarization marker of M2 microglia). Real-time polymerase chain reaction was employed to determine the mRNA levels of iNOS, CD206, pro-inflammatory cytokines [tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6] and anti-inflammatory cytokines (IL-4 and IL-10) in the hippocampus. Western blotting was employed to determine the protein levels of iNOS and CD206 in the hippocampus. The levels of IL-4 and IL-6 in the hippocampus were detected by enzyme-linked immunosorbent assay. ResultCompared with the normal control group, the model rats showed a reduction in sucrose preference (P<0.05), an increase in immobility time (P<0.05), decreased motor and exploratory behaviors (P<0.05), and weakened learning and spatial memory (P<0.05). In addition, the model rats showed up-regulated mRNA and protein levels of iNOS and mRNA levels of IL-1β, IL-6, and TNF-α (P<0.05). Compared with the model group, Baihuan Xiaoyao decoction increased the sucrose preference value (P<0.05), shortened the immobility time (P<0.01), increased the motor and exploratory behaviors (P<0.05), and improved the learning and spatial memory (P<0.01). Furthermore, the decoction down-regulated the positive expression and protein level of iNOS, lowered the levels of TNF-α, IL-1β, and IL-6 (P<0.01), promoted the positive expression of CD206, and elevated the levels of IL-4 and IL-10 (P<0.01) in the hippocampus of the high dose group. Moreover, the high-dose Baihuan Xiaoyao decoction group had higher sucrose preference value (P<0.01), shorter immobility time (P<0.01), longer central distance (P<0.01), stronger learning and spatial memory (P<0.01), higher positive expression and protein level of iNOS (P<0.01), lower levels of TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01), lower positive expression and mRNA level of iNOS (P<0.05), and higher levels of IL-4 and IL-10 (P<0.05, P<0.01) than the fluoxetine group. ConclusionBaihuan Xiaoyao decoction can improve the depression-like behavior of juvenile rats by inhibiting the M1 polarization and promoting the M2 polarization of microglia in the hippocampus.