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Objective To investigate the expression of circRAF1 in primary ovarian insufficiency(POI)and explore its effect on cell proliferation and apoptosis of human ovarian granulosa cells(GCs)line(KGN cells).Methods The expression of circRAF1 in GCs and serum of patients with normal ovarian reserve function(n = 50)and patients with POI(n = 50)were detected with RT-qPCR.The correlation of circRAF1 with ovarian reserve function indexes was analyzed.Small interfering RNA(siRNA)targeting circRAF1 was constructed and trans-fected into KGN cells,with the cell proliferation detected by CCK-8 and EdU assay,and the cell apoptosis detected by JC-1 and Tunel assay.The mRNA and protein levels of genes related to cell proliferation and apoptosis(FSHR,PCNA,Bcl-2,Casp-9,Bax)were detected by RT-qPCR and WB.Results The expression of circRAF1 decreased in GCs and serum of POI patients.The expression of circRAF1 was positively correlated with serum E2 and AMH levels(P<0.001),but negatively correlated with serum FSH and LH levels(P<0.001).At the same time,the expression of circRAF1 was positively correlated with AFC(P<0.001).Interfering with the expression of circRAF1 could inhibit the proliferation of KGN cells and promote their apoptosis.Conclusion The expression of circRAF1 in the GCs and serum of POI patients is down-regulated,which is correlated with the decline of ovarian reserve function.Interfering with circRAF1 can inhibit the proliferation of GCs and promote their apoptosis.
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BACKGROUND:Previous studies have shown that silent mating-type information regulator 2 homolog 1(SIRT1)-mechanistic target of rapamycin(mTOR)signaling plays an important role in the progression of osteoarthritis.Emodin has a protective effect on osteoarthritic chondrocytes. OBJECTIVE:To investigate the effects of emodin on the proliferation,apoptosis and oxidative stress of osteoarthritic chondrocytes based on the SIRT1-mTOR signaling. METHODS:Rat chondrocytes were isolated and cultured in vitro.Osteoarthritic chondrocyte model in vitro was induced by 10 ng/mL interleukin-1β.Cell counting kit-8 method was used to determine the viability of rat chondrocytes treated with 0,20,40,80,120,160 μmol/L emodin,and the appropriate concentration of emodin was selected.Rat chondrocytes isolated and cultured in vitro were randomly divided into control group,model group,low-dose emodin group,high-dose emodin group,EX527 group,and high-dose emodin+EX527 group.In vitro osteoarthritis model was constructed by induction of 10 ng/mL interleukin 1β in all groups except the control group.The cells in the latter four groups were correspondingly treated with emodin or/and EX527.The proliferation and apoptosis of chondrocytes in each group were detected by cell counting kit-8,Edu staining and flow cytometry respectively.The relative content of reactive oxygen species and the levels of malondialdehyde,superoxide dismutase,catalase,and glutathione peroxidase in chondrocytes of rats in each group were measured with the kit.The expression of proteins related to cell matrix degradation,apoptosis and the SIRT1-mTOR pathway-related proteins in each group were detected by western blot. RESULTS AND CONCLUSION:Compared with the control group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the model group were decreased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were increased(P<0.05).Compared with the model group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the low-and high-dose emodin groups were increased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were decreased(P<0.05).Compared with the low-and high-dose emodin groups,the indexes of EX527 group showed the opposite trend(P<0.05).Compared with the high-dose emodin group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the high-dose emodin+EX527 group were decreased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were increased(P<0.05).To conclude,emodin can inhibit oxidative stress of osteoarthritic chondrocytes by activating the SIRT1-mTOR signaling,thereby promoting chondrocyte proliferation and reducing apoptosis.
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According to different biological functions, breast cancer can be divided into triple-negative breast cancer and non-triple-negative breast cancer. The former has stronger proliferation and invasion ability and faster metastasis. Its immune tissue indicators are estrogen receptor (estrogen receptor, ER) and progesterone receptor. Both the body (progesterone receptor,PR) and human epidermal growth factor 2 receptor (human epidermal growthFactor receptor, HER2) are negative. Surgery and radiotherapy are traditional therapies for TNBC, but these therapies have many side effects and poor prognosis. In order to better prevent the spread and treatment of TNBC, it is particularly important to study the growth, proliferation and apoptosis mechanisms of cancer cells. In recent years, studies have shown that a variety of signaling pathways, lncRNA, and miRNA all affect the growth process of TNBC. In recent years, studies have shown that a variety of signaling pathways, lncRNA, and miRNA all affect the growth process of TNBC. This article reviews domestic and foreign studies on the relevant regulatory mechanisms of TNBC cell proliferation and apoptosis, aiming to clarify the importance of breast cancer cell proliferation and apoptosis pathways, and provide new ideas for drug development for TNBC targeted therapy.
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Objective:To study the effect of icaritin on the proliferation， apoptosis， migration and invasion of human epithelial ovarian cancer A2780 cells and the inhibitory mechanism of icaritin against cell invasion and migration via the regulation of epithelial-mesenchymal transformation （EMT）-related molecule expression. Method:A2780 cells were divided into the blank control group and low-， medium-， and high-dose （5， 10， 20 μmol·L<sup>-1</sup>） icaritin groups and received the corresponding inventions for 48 h. Cell proliferation and viability were detected using the cell counting kit-8 （CCK-8）. The cellular proliferation inhibition and apoptosis rates were assayed by flow cytometry. The cell invasion and migration were observed in Scratch test and transwell test， followed by the calculation of wound healing rate and migration rate. The protein and mRNA expression levels of EMT-related molecules including E-cadherin， N-cadherin， and Vimentin and tumor invasion and migration-related molecule matrix metalloproteinase-9 （MMP-9） were measured by Western blot and real-time polymerase chain reaction （Real-time PCR）. Result:As revealed by CCK-8 assay and flow cytometry， compared with the blank control group， the icaritin groups all exhibited elevated proliferation inhibition rate （<italic>P</italic><0.01） and apoptosis rate （<italic>P</italic><0.05）. According to the Scratch test and transwell test， compared with the blank control group， the icaritin groups displayed weakened invasion and migration ability and decreased number and rate of cell invasion and migration （<italic>P</italic><0.05）. Western blot and Real-time PCR results showed that the protein and mRNA expression levels of N-cadherin， MMP-9 and Vimentin in each icaritin group were down-regulated as compared with those in the blank control group， while the expression of E-cadherin was up-regulated. Conclusion:Icaritin inhibits the proliferation and promotes the apoptosis of human ovarian cancer A2780 cells， and it inhibits the invasion and migration of A2780 cells possibly by regulating the expression of EMT-related molecules.
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To investigate the expression of AGAP2-AS1 in colon cancer, its influence on the malignant behavior of colon cancer cells, and its effects on the Yes-associated protein (YAP) signaling pathway. Methods: The expression of AGAP2-AS1 in 457 colon cancer samples and 42 healthy samples was obtained from TCGA. Twenty matched colon cancer and adjacent cancer tissues were collected from patients diagnosed in Yiwu Central Hospital and confirmed by pathology department from January 2018 to March 2019. The expression of AGAP2-AS1 in these tissues, as well as in SW480, HCT-116, and NCM460 cells, was detected by real-time fluorescent quantitative PCR. The changes in cell proliferation, migration and apoptosis were analyzed and detected using a CCK-8 kit, and clone formation, wound-healing assay, and flow cytometry experiments. The levels of YAP, p-YAP, matrix metalloproteinase-2 (MMP2), and MMP9 were evaluated by Western blot. Immunofluorescence was used to examine the localization of YAP in HCT-116 cells. The levels of YAP, p-YAP, MMP2, and MMP9 proteins were evaluated after the co-transfection of pcDNA3.1-AGAP2-AS1 and si-YAP plasmids. Results: TCGA indicated that the expression of AGAP2-AS1 was significantly increased in colon cancer tissues. Similarly, in the present study, AGAP2-AS1 expression was significantly increased in colon cancer tissues and cells. After AGAP2-AS1 knockdown/overexpression, the capacity of migration and proliferation was significantly reduced/increased, and apoptosis was significantly increased/inhibited in colon cancer cells. Moreover, AGAP2-AS1 knockdown triggered the phosphorylation of YAP, and a significant reduction in the expression of MMP2 and MMP9 (P<0.05). Co-transfection studies revealed that AGAP2-AS1 upregulated the expression of MMP2 and MMP9 via activation of the YAP pathway (P<0.05). Conclusions: AGAP2-AS1 was highly expressed in colon cancer tissues and cell lines. AGAP2- AS1 can induce proliferation and migration, and inhibit apoptosis in colon cancer cells. In addition, AGAP2- AS1 regulated MMP2 and MMP9 expression by activating the YAP pathway, thereby affecting colon cancer metastasis.
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Objective To investigate the possible molecular mechanism for alpha(α)-mangostin's inhibition of the proliferation and apoptosis of human gastric cancer cells.Methods Human gastric adenocarcinoma SGC7901 cell line was treated with α-mangostin.CCK8 method was used to measure the viability of SGC7901 cells.The effect of α-mangostin on apoptosis and cell cycle was determined by immune fluorescence and flow cytometry.The expression of the relevant proteins was detected using Western blot.The shapiro-wilk test was performed for evaluation of deviation from normality.Normally distributed data was analyzed with one-way ANOVA.Welch test was used in data with heterogeneity of variance and multiple compared by Games-Howell test after that.Results CCK8 results showed that cell viability differed significantly among groups treated with different concentrations of α-mangostin(10,15,20,25,and 30 μmol/L)(P＜0.05).QPCR data showed that the concentration of α-mangostin was positively correlated with mRNA level of LC 3 but not caspase protein(r=0.976,P＜0.05).In 15 μmol/L but not 10 μmol/L α-mangostin treatment system,the autophagy inhibitors 3-MA(10 μmol/L),bafilomycin A(10 μmol/L)and LY294002(10 μmol/L)could significantly alleviate α-mangostin's killing effect on SGC7901 cells(P＜0.05).Conclusion The anti-tumor effects of α-mangostin against human gastric adenocarcinoma cells in vitro can be partly attributed to apoptosis,autophagy and arresting cell phase.
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Gastric cancer is a common highly aggressive malignant neoplasm .Studies have found that exosomes can carry a va-riety of biologically active components in multiple cells , such as various proteins , dsDNA and microRNAs .Specific membrane structure and contents of exosomes are widely involved in material exchange between gastric cancer cells , such as the formation of gastric cancer microenvironment , promoting gastric cancer cell proliferation and apoptosis , invasion and metastasis , tumor resistance and so on .Re-search on exosomes may be a breakthrough in finding new ways to treat gastric cancer .In this paper , we discuss the research progress of exosomes in gastric cancer .
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Gastric cancer is a common highly aggressive malignant neoplasm .Studies have found that exosomes can carry a va-riety of biologically active components in multiple cells , such as various proteins , dsDNA and microRNAs .Specific membrane structure and contents of exosomes are widely involved in material exchange between gastric cancer cells , such as the formation of gastric cancer microenvironment , promoting gastric cancer cell proliferation and apoptosis , invasion and metastasis , tumor resistance and so on .Re-search on exosomes may be a breakthrough in finding new ways to treat gastric cancer .In this paper , we discuss the research progress of exosomes in gastric cancer .
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To explore the effect of leech on proliferation and apoptosis of vascular smooth muscle cells(VSMCs) in early atherosclerosis rats via p38MAPK signaling pathway and investigate its possible mechanism. Biochemical analyzer was used to examine the regulation of leech on levels of triglycerides(TG), total cholesterol(TC), low-density lipoprotein(LDL-C), and high-density lipoprotein(HDL-C) in blood lipid of rats. The expression of transforming growth factor-beta 1(TGF-β1) in serum was detected by ELISA. Immunological histological chemistry (IHC) was taken to measure the expression levels of proliferating cell nucleus antigen(PCNA) and cell apoptosis proteinase-3(Caspase-3), while the protein expression levels of MKK3, p38 and C-myc were detected by Western blot. In addition, hematoxylin and eosin(HE) staining was used to observe the morphological change of thoracic aortas. The results showed that leech decreased the levels of TC, LDL-C obviously and increased HDL-C, suppressed the expression levels of TGF-β1 and PCNA, up-regulated Caspase-3, down-regulated the expression levels of MKK3, p38, and C-myc protein. HE staining indicated that it could inhibit intimal thickening and reduce plaque formation. The above results indicated that leech may affect the protein expression of the p38MAPK signaling pathway to inhibit proliferation and promote the apoptosis of VSMCs via reducing blood lipid levels and suppressing TGF-β1, aiming at inhibiting intimal thickening and reducing plaque formation, tand then slowing down the process of early atherosclerosis.
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Objective To discuss the influence of different concentration sulindac on pancreatic cancer cell line PANC-1 cell proliferation and apoptosis,and investigate the possible mechanism that sulindac can inhibit the Wnt/beta-catenin pathway to kill pancreatic cancer cells. Methods PANC-1 cell were divided into negative control group (added containing no sulindac DMSO)and experimental group (added sulindac with concentrations of 0.25 ,0.5 ,1 ,1.5 ,2 mM medium,respectively,name as 0.25 mM group,0.5 mM group,1.0 mM group,1.5 mM group,2.0 mM group),and treated with different time,cell proliferation inhibition ratio in each group was detected by MTT assay,cell apoptosis ratio was detected by flow cytometry,the expression ofβ-catenin mRNA and protein were detected by RT-PCR and immunocytochemistry.Results MTT results showed that sulindac can inhibit the cell proliferation of PANC-1 by a dose-and time-dependent manner.Cell apoptosis increased after sulindac treatment in different degrees,and there were statistical differences between 1.5,2.0 mMgroup and control groups (P<0.05).RT-PCR results showed that the expression ofβ-catenin mRNA decreased after the treatment of sulindac,there were statistical differences between 1.5,2.0 mMgroup and control group (P<0.05). In the 2.0mM group,the expression ofβ-catenin decreased along with the time extending (P<0.05 ).ICC results showed that sulindac inhibited the expression ofβ-catenin protein and nuclear accumulation,there were no statistical differences in 0.25 ,0.5 mM group and control group,but there were statistical differences in 1.0,1.5,2.0 mMgroup.Conclusion Sulindac could inhibit cell proliferation and facilitate apoptosis of PANC-1,this effect is dose-and time-dependent.The inhibition of Wnt/beta-catenin signal pathway may be a possible mechanism of its cytotoxicity.
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Emodin has been known for its various pharmacological functions, including anti-rumor, anti-inflanmation,the inhibition of cell proliferation and secretion of pancreatic enzyme, acceleration of osteoblast cell differentiation and fibrinolysis, and promotion of wound healing. The researching progress of anti-tumor function of emodin was reviewed in this paper for the purpose of understanding it's mechanism of anti-tumor comprehensively.