Effect and mechanism of atorvastatin on the proliferation，autophagy and glucose metabolism of AGS cells in human gastric cancer / 中国药房

OBJECTIVE To explore the effects and mechanism of atorvastatin on the proliferation， autophagy and glucose metabolism of AGS cells in human gastric cancer. METHODS The effects of low， medium and high concentrations of atorvastatin （12.5， 25， 50 μmol/L） on the viability of AGS cells were investigated through preliminary experiments， and the concentration of action was screened. The formal experiment was divided into control group （no intervention）， atorvastatin group （25 μmol/L）， positive control group （50 mg/L 5-fluorouracil）， inhibitor group [25 μmol/L atorvastatin +10 μmol/L phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt） signaling pathway inhibitor LY294002] and activator group （25 μmol/L atorvastatin +10 μmol/L PI3K/Akt signaling pathway activator SC79）， all of which were treated for 24 h. Glucose metabolism （glucose and lactic acid contents） and cell proliferation rate were detected， as well as the expression of autophagy-associated protein light chain 3 （LC3） Ⅰ， LC3Ⅱ and PI3K/Akt signaling pathway-associated proteins in cells. RESULTS Both medium and high concentrations of atorvastatin could significantly inhibit the viability of AGS cells （P＜0.05）， and 25 μmol/L atorvastatin was selected for the official experiment for follow-up experiments. Compared with the control group， the contents of glucose and lactic acid， cell proliferation rate， p-PI3K/PI3K and p-Akt/Akt ratios in the positive control group and atorvastatin group were significantly decreased （P＜ 0.05）， and the protein expression levels of LC3 Ⅰ and LC3 Ⅱ were significantly increased （P＜0.05）. Compared with the atorvastatin group， the inhibitor further promoted the changes in the above indexes （P＜0.05）， and the activator significantly reversed the changes in the above indexes （P＜0.05）. CONCLUSIONS Atorvastatin could inhibit glucose metabolism and proliferation of AGS cells in human gastric cancer and promote autophagy. The mechanism may be related to the inhibition of the PI3K/Akt signaling pathway.

Effects of gypenosides on proliferation and apoptosis of human gastric cancer cells and its mechanism / 中国药理学通报

Aim To explore the effect of gypenosides on proliferation and apoptosis of human gastric cancer cells SGC-7901 and AGS and its mechanism. Methods Different concentrations of gypenosides were cultured with human gastric cancer cells SGC-7901 and AGS. Cell viability assay was used to detect cell proliferation activity, and the IC

Effect of Circular RNA hsa_circ_0067582 on the Proliferation and Invasion Ability of Gastric Cancer Cells / 中国医学科学院学报

Objective To investigate the effects on cell proliferation and invasion of the circular RNA hsa_circ_0067582 in gastric cancer(GC). Methods After hsa_circ_0067582 overexpression (Oe-circ_0067582) plasmid was transfected into AGS and SGC-7901 cells,the cell viability,proliferation,invasion ability,and apoptosis were detected by CCK-8,colony formation and EdU assays,Transwell assay,and flow cytometry,respectively.Western blotting was employed to detect the expression levels of proteins related to the cell apoptosis and epithelial-mesenchymal transition(EMT).The effect of Oe-circ_0067582 on the growth of SGC-7901 cells in nude mice was observed.Bioinformatics tools were used to predict the binding target miRNA of hsa_circ_0067582,and the competing endogenous RNA(ceRNA)regulatory network was established.Finally,functional enrichment was performed to analyze the biological functions of the target genes of the predicted miRNA. Results Compared with the pLO-ciR(empty plasmid)group,the Oe-circ_0067582 group in AGS and SGC-7901 cells attenuated the cell viability(t=7.883,P=0.001;t=5.679,P=0.005),proliferation(t=6.709,P=0.003;t=5.857,P=0.003),and invasion ability(t=7.782,P=0.002;t=6.342,P=0.003)and induced cell apoptosis(t=7.225,P=0.002;t=11.509,P=0.001).Western blotting showed that the Oe-circ_0067582 group in AGS and SGC-7901 cells up-regulated the protein levels of cysteinyl aspartate specific proteinase (Caspase) 3(t=6.863,P=0.002;t=7.024,P=0.001),Caspase 7(t=3.295,P=0.04;t=6.008,P=0.004),Caspase 9(t=4.408,P=0.012;t=6.278,P=0.004),and E-cadherin(t=12.453,P=0.002;t=10.867,P=0.001),while down-regulated those of Vimentin(t=7.242,P=0.002;t=5.694,P=0.004)and N-cadherin(t=6.480,P=0.003;t=7.446,P=0.001).Furthermore,Oe-circ_0067582 significantly inhibited the growth of tumor in the SGC-7901 tumor-bearing nude mice(t=3.526,P=0.017).The prediction based on TargetScan and miRnada suggested that hsa_circ_0067582 can competitively bind to hsa-miR-181b-3p,hsa-miR-337-3p,hsa-miR-421,and hsa-miR-548d-3p.The functional enrichment indicated that the target genes of miRNA were involved in multiple cancer-related biological processes including negative regulation of apoptotic process,gene expression,transcriptional misregulation in cancer,transforming growth factor-β,and p53 signaling pathways. Conclusion Oe-circ_0067582 can inhibit the proliferation and attenuate EMT process to reduce the invasion ability of AGS and SGC-7901 cells,which provides a new target for the treatment of GC.

Expression of mitogen-activated protein kinase kinase 1 in gastric cancer and its clinical significance / 解剖学报

Objective To investigate the expression of mitogen-activated protein kinase kinase 1 (MAP2K1) in gastric cancer and its clinical significance. Methods Immunohistochemistry and Western blotting were used to detect the protein expression of MAP2K1 in gastric cancertissues and cells. The morphology and the expression position of MAP2K1 were observed by immunofluorescence. MAP2K1 mRNA expression in gastric cancer tissues was analyzed by data mining of Starbase database and Oneomine database. The correlation between MAP2K1 mRNA expression and clinicopathological features was analyzed by UALCAN database. Survival analysis was performed using Kaplan Meier-Plotter online analysis tools. GEPIA2 database mining the relationship between MAP2K1 and gastric cancer stem cell related factors and drug resistance related factors. Results Immunohistochemistry, immunofluorescence and Western blotting showed that MAP2K1 protein was highly expressed in gastric cancer tissues and cells, and MAP2K1 was expressed in the cytoplasm of gastric cancer. According to the analysis of various databases, the expression of MAP2K1 mRNA in gastric cancer tissue was higher than that in normal gastric tissue, and the expression of MAP2K1 mRNA was closely related to gastric cancer stage, grade, lymph node metastasis and patient gender, and the overall survival rate of gastric cancer patients in the group with high MAP2K1 mRNA expression was significantly lower than that in the group with low MAP2K1 mRNA expression, which may be related to the characteristics of gastric cancer stem cells and drug resistance. Conclusion MAP2K1 is highly expressed in gastric cancer, and its expression level may affect the poor prognosis of patients by regulating stem cell related factors and drug resistance related factors. MAP2K1 may be a new diagnostic marker to determine the prognosis of gastric cancer patients.

Extract from Zanthoxylum piperitum Induces Apoptosis of AGS Gastric Cancer Cells Through Akt/MDM2/p53 Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To determine the effect of Zanthoxylum piperitum extracet (ZPE) on apoptosis and analyze anticancer substances in ZPE, changes in proteins related to apoptosis, and pathological changes in tumors in mouse.@*METHODS@#Fifteen 4-week-old female BALB/c nu/nu mice were divided into 3 groups depending on ZPE dose, with 5 in each group. AGS gastric carcinoma cells (1 × 10@*RESULTS@#High performance liquid chromatography (HPLC) analysis showed that ZPE contained organic sulfur compounds such as alliin and S-allylcysteine. MTT assay results revealed that ZPE (10-85 µ g/mL) could effectively inhibit the growth of AGS gastric cancer cells at higher concentrations (P<0.05, P<0.01). The annexin V & dead cell staining assay and cell cycle arrest assay confirmed a dose-dependent increase in the apoptosis rate and G@*CONCLUSION@#ZPE decreases AGS cell proliferation and induces apoptosis by inhibiting Akt and MDM2 expression.

Curcuma wenyujin Y. H. Chen et C. Ling n-Butyl Alcohol Extract Inhibits AGS Cell Helicobacter pylori Promoted Invasiveness by Down-Regulating Caudal Type Homeobox Transcription Factor and Claudin-2 Expression / 中国结合医学杂志

OBJECTIVE@#To investigate the effects and possible mechanisms of action of Curcuma wenyujin Y. H. Chen et C. Ling n-Butyl alcohol extract (CWNAE) on repression of human gastric cancer (GC) AGS cell invasion induced by co-culturing with Helicobacter pylori (HP).@*METHODS@#AGS cells were cultured with HP of positive or negative cytotoxin-associated gene A (CagA) and vacuolating cytotoxin gene A (VacA) expression (CagA+/- or VacA+/-) and divided into 5 group. Group A was cultured without HP as a control, Group B with HP, Group C with HP, Group D with HP and CWNAE, and Group E with HP and CWNAE. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) and tumor invasion assays, examinations of morphology and ultramicroscopic structures, quantitative real-time polymerase chain reaction and Western blots were performed to measure the effects and uncover the mechanisms behind these effects of HP and CWNAE on the epithelial-mesenchymal transition (EMT) of AGS cells.@*RESULTS@#The 10% inhibitory concentration of CWNAE against AGS cells after a 48 h incubation was 19.73±1.30 μg/mL. More AGS cells were elongated after co-culturing with HP than after culturing with HP. In tumor invasion assays, HP significantly enhanced the invasiveness of AGS cells compared to the other experimental groups (all P value <0.05), and this effect was inhibited by CWNAE. Treatment with CWNAE normalized tight junctions and reduced the number of pseudopodia of AGS cells co-cultured with HP. HP up-regulated zincfinger ebox binding homeobox 1 (ZEB1) in AGS cells after co-culturing for 24 h. Expression of caudal type homeobox transcription factor (CDX-2) and claudin-2 was significantly increased by HP (P<0.05), but not by HP.@*CONCLUSION@#HP promoted the invasiveness of AGS cells through up-regulation of ZEB1 transcription and claudin-2 and CDX-2 expression. CWNAE inhibited these effects of HP on AGS cells by down-regulating ZEB1 transcription, and CDX-2 and claudin-2 expression.

lncRNA MAFG-AS1 promotes migration, invasion and aerobic glycolysis of gastric cancer AGS cells by regulating miR-11181-3p/GLG1 axis / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of lncRNA MAFG-AS1/ miR-11181-3p/GLG1 axis on cell migration, invasion and aerobic glycolysis of gastric cancer (GC) cells and its possible mechanism. Methods: AGS, a GC cell line with relatively high expression of MAFG-AS1, was selected as the study object. qPCR was used to detect RNA expression levels of MAFG-AS1, miR-11181-3p and GLG1. Transwell and glycolysis analysis were used to investigate cell migration, invasion and aerobic glycolysis. Bioinformatics analysis and Dual luciferase reporter gene assay were used to analyze the interaction among MAFG-AS1, miR-11181-3p and GLG1. Results: Knockdown of MAFG-AS1 significantly up-regulated miR-11181-3p and down-regulated GLG1 expression (both P<0.01), and significantly inhibited migration, invasion and aerobic glycolysis of GC cells (all P<0.01). Luciferase reporter gene assay confirmed that MAFG-AS1 competitively sponged miR-11181-3p (P<0.01). Inhibition of miR-11181-3p or overexpression of GLG1 partially reversed the inhibitory effect of MAFG-AS1 knockdown on GC cell migration, invasion, and aerobic glycolysis (all P<0.05 or P<0.01). Conclusion: MAFG-AS1 promotes cell migration, invasion and aerobic glycolysis of GC cells via miR-11181-3p/GLG1 axis, and may be a potential molecular target for GC diagnosis and therapy.

lncRNA XIST promotes gastric cancer progression via regulating miR-337-3p/ HOXC8 axis / 中国肿瘤生物治疗杂志

@# Objective: To investigate the mechanism of lncRNA XIST (XIST) on modulating gastric cancer progression via regulating miR-337-3p/HOXC8 axis. Methods: A total of 58 cases of gastric cancer tissues and corresponding para-cancerous tissues resected from March 2013 to January 2018 in Department of General Surgery, Kailuan General Hospital of Tangshan City were collected for this study; in addition, human gastric cancer cell lines (AGS, MGC803, HGC27) and human gastric mucosal GES-1 cells were also collected. qPCR was used to detect the expressions of XIST and miR-337-3p in above mentioned gastric tissues and cell lines. XIST-knockdown vectors, miR-337-3p mimics, miR-337-3p inhibitor and HOXC8-overexpression vectors were transfected into AGS cells. The proliferation and invasion of AGS cells were detected by CCK-8 and Transwell experiments respectively, and the expression levels of HOXC8, E-cadherin, N-cadherin and vimentin were detected by WB. The targeting relationships between XIST, miR337-3p and HOXC8 were verified by dual-luciferase reporter gene assay. Results: XIST was up-regulated in gastric cancer tissues and cell lines (all P<0.01). XIST knockdown significantly inhibited proliferation, invasion and EMT of AGS cells (P<0.05 or P<0.01). Moreover, XIST directly interacted with miR-337-3p and down-regulated its expression, while HOXC8 was the target gene of miR-3373p. Furthermore, XIST knockdown suppressed proliferation, invasion and EMT ofAGS cells through up-regulating the inhibitory effect of miR-337-3p on HOXC8 (P<0.05 or P<0.01). Conclusion: XIST knockdown can suppress the proliferation, invasion and EMT of AGS cells, which may be related with down-regulation of HOXC8 by targeting miR-337-3p.

Effect of DKK1 on proliferation, cell cycle and apoptosis of gastric cancer AGS cells / 中国肿瘤生物治疗杂志

@# Objective： ：To study the effect of silencing DKK1 (Dickkopf1) gene on the proliferation, cell cycle and apoptosis of gastric cancer AGS cells and the action mechanism. Methods： ：The DKK1-shRNA vector was constructed and transfected into AGS cells. The stably transfected cell lines were screened. The total protein and RNAof the transfected cells were extracted and the mRNAand protein expressions of DKK1 were detected by qPCR and WB, respectively. The experiment was divided into blank control group (Control), negative control group (shNC) and DKK1 silence group (DKK1-shRNA). CCK8 assay was used to detect the proliferation ofAGS cells of each group cultured for 0, 24, 48, 72, 96, 120 and 144 h, and flow cytometry was used to analyze the cell cycle and apoptosis in each group. The relationship between DKK1 and clinicopathological features of gastric cancer was analyzed after searching HPA database. Results：The gastric cancer AGS cells with stable DKK1 gene knockdown was successfully established, and it was confirmed that the mRNA and proteinexpressions of DKK1 in DKK1-shRNA group decreased by 72% and 47%, respectively, compared to shNC group (all P<0.05). The cell proliferation curve showed that, the cell proliferation in DKKl-shRNAgroup significantly decreased after 72 hour of culture compared with that in control and shNC groups (P<0.05). The cell number of S phase decreased from 32.06% to 25.87%, while the number of G2/M phase increased from 8.49% to 21.26% compared with shNC group (all P<0.05). The number of apoptotic cells also statistically increased from 10.34% to 20.65% (all P<0.05). The data of HPAdatabase showed that DKK1 mRNAlevel in gastric cancer tissues was significantly higher than that in normal tissues, and the high expression of DKK1 mRNAwas negatively correlat ed with the survival rate of gastric cancer patients. Conclusion: : Silencing DKK1 gene can inhibit the proliferation of gastric cancer cells, arrest cells in G2/M phase and promote cell apoptosis. DKK1 plays a pro-carcinogenic effect in gastric cancer.

Effects of hesperidin on apoptosis of human gastric cancer AGS cells and its mechanism / 中草药

Objective: To investigate the effect of hesperidin on apoptosis of gastric cancer AGS cells and its related molecular mechanisms. Methods: MTT assay was used for the killing effect of hesperidin on human gastric cancer AGS cells; Annexin V-FITC/PI double staining and flow cytometry was used to detect the apoptosis induced by hesperidin on AGS cells, the level of reactive oxygen species, and the addition of NAC Post-apoptosis changes; Western blotting was used to detect the expression of apoptosis-related proteins and signaling pathway-related proteins. Results: MTT assay showed that hesperidin had a good inhibiting effect on AGS cells. After treated with hesperidin, AGS cells showed apoptosis such as nuclear condensation and cell shrinkage. Annexin V-FITC/PI double staining and flow cytometry showed that hesperidin can induce mitochondrial dependent apoptosis of AGS cells and increase the level of intracellular reactive oxygen species. After pretreatment of NAC, hesperidin induced apoptosis inhibition. The results of Western blotting showed that the expression of p-JNK, p-p38, Bad, cleaved Caspase-3, and cleaved PARP increased, and the expression of anti-apoptotic proteins p-ERK and Bcl-2 decreased, which indicated that hesperidin activated the MAPK signaling pathway and mitochondria-dependent apoptosis in AGS cells. Conclusion: Hesperidin has a good killing effect on human gastric cancer AGS cells, and induces mitochondria-dependent apoptosis in AGS cells by increasing the level of reactive oxygen species in AGS cells and regulating MAPK signaling pathway.

In vitro and in vivo inhibition of Helicobacter pylori by Lactobacilllus paracasei HP7 / 한국실험동물학회지

The efficacy of standard therapeutic strategies for Helicobacter pylori (H. pylori) infection is decreasing over time due to the emergence of drug-resistant strains. As an alternative, the present study investigated the capacity of Lactobacilllus paracasei (L. paracasei) HP7, isolated from kimchi, to inhibit H. pylori growth. The effects of L. paracasei HP7 on H. pylori adhesion and H. pylori-induced inflammation were examined in AGS human gastric adenocarcinoma epithelial cells and a mouse model of H. pylori SS1 infection. L. paracasei HP7 reduced H. pylori adhesion to AGS cells and suppressed the inflammatory response in infected cells by downregulating interleukin-8. H. pylori colonization in the stomach of C57BL/6 mice was demonstrated by rapid urease test, and results showed significant decrease in mice post-treated with L. paracasei HP7. Additionally, L. paracasei HP7 decreased gastric inflammation and epithelial lesions in the stomach of H. pylori-infected mice. These results demonstrate that L. paracasei HP7 treatment can inhibit H. pylori growth and is thus a promising treatment for patients with gastric symptoms such as gastritis that are caused by H. pylori infection.

Expression of FMN2 in gastric adenocarcinoma tissues and its clinical significance / 中国肿瘤生物治疗杂志

@# Objective: To investigate the expression of Formin-2（FMN2）protein in gastric cancer tissues and its correlation to clinicopathological features of gastric cancer patients, as well to explore its effect on the proliferation of gastric adenocarcinoma AGS cells. Methods: 84 cases of gastric adenocarcinoma tissues and corresponding para-cancerous tissues were surgically collected from patients treated in the First Affiliated Hospital of Air Force Military Medical University from September 2015 to September 2017. The expression of FMN2 in gastric adenocarcinoma tissues was detected by immunohistochemical staining and analyzed with RNA-Seq data-sets GEPIA. The relationship between FMN2 protein expression in gastric adenocarcinoma tissues and its clinicopathological features was also explored. MTT assay was used to detect the effect of FMN2 onAGS cell proliferation activity, and Western blotting was used to detect the effect of FMN2 on the expression of apoptosis-related protein caspase-3 in AGS cells. Results: The expression level of FMN2 in gastric adenocarcinoma tissues was significantly lower than that in matched adjacent tissues and the expression level of FMN2 was closely related to the TNM stage and differentiation of gastric adenocarcinoma (all P<0.05). Compared toAGS control group, the proliferation activity of AGS/FMN2 was significantly decreased and the expression of apoptosis-related gene Caspase-3 was markedly increased (all P<0.05). Conclusion: The expression of FMN2 was significantly decreased in gastric adenocarcinoma tissues and its low expression is closely related to the degree of tumor differentiation and clinical TNM stage. Moreover, FMN2 over-expression significantly decreased the proliferation of AGS cells. FMN2 may function as independent risk factor for the prognosis of gastric adenocarcinoma, which may provide new ideas for the treatment of gastric adenocarcinoma.

Correlation between ATF5 protein expression level and drug sensitivity of gastric cancer AGS cells / 实用医学杂志

Objective To explore the correlation between ATF5 protein expression level and drug sensitivity of gastric cancer AGS cells.Methods We established the high,middle and low ATF5 expression groups by transfecting the AGS cells with ATF5 pcDNA3.1 plasmid,ATF5 siRNA and empty plasmid using the liposome transfection technique.Then we detected the expression of the ATF5 protein of the 3 groups using the Western Blot.The drug sensitivity of AGS cells in 3 groups to paclitaxel and cisplatin was detected using the MTT experiments and plate clone formation experiments then the correlation between ATF5 protein expression level and drug sensitivity of gastric cancer AGS cells was analyzed.Results With the effect of paclitaxel or cisplatin on AGS cells in 3 groups,the surviving fraction and the IC50 of high ATF5 expression group were higher than those of control group,which indicated statistical significance (all P ＜ 0.05);while the surviving fraction and the IC50 of low ATF5 expression group were lower than those of control group (all P ＜ 0.05,P cisplatin =0.00,P paclitaxel =0.002).We found that cell clone formation of AGS cells to paclitaxel and cisplatin was significantly increased after the transfection with ATF5 protein plasmid and decreased after transfection with ATF5 siRNA plasmid (all P ＜ 0.05).Conclusion The expression of ATF5 protein is related to the drug sensitivity of gastric cancer AGS cells that upregulation of ATF5 protein expression can decrease the drug sensitivity of AGS cells to paclitaxel and cisplatin in gastric cancer.

Inhibition of annonaceous acetogenins on cell activities of human gastric cancer cells in vitro / 中草药

Objective To study the effect of annonaceous acetogenins (ACGs) on human gastric cancer cells in vitro. Methods After ACGs were administered to gastric cancer cells in vitro, the cell viability, cell adhesion ability and cell migration ability were assessed by MTT assay, adhesion assay and wound-healing assay, respectively. Results ACGs inhibited the cell viability, adhesion ability and migration ability in a dose-dependent manner in gastric cancer cells. Conclusion ACGs could inhibit cell activities of human gastric cancer cells in viro, and will be developed as a promising anticancer candidate and used in gastric cancer.

Anti-Helicobacter and Anti-inflammatory Effects of Sohamhyungtang in Helicobacter pylori-Infected Human Gastric Epithelial AGS cells

This study evaluated the anti-Helicobacter and anti-inflammatory effects of Sohamhyungtang (SHHT). The minimum inhibitory concentration (MIC) of SHHT against Helicobacter pylori (H. pylori) was determined by the agar dilution method. Expression of the H. pylori cagA gene in the presence of SHHT was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Inhibition of H. pylori urease by SHHT was determined by the phenol-hypochlorite assay. Antiadhesion activity of SHHT was measured by ureaphenol red reagent. Inhibition of nitric oxide (NO) production in AGS cells was measured with Griess reagent. Inducible nitric oxide synthase (iNOS) and IL-8 mRNA expression in AGS cells which were infected with H. pylori was determined by qRT-PCR. IL-8 level was measured by enzyme-linked immunosorbent assay (ELISA). The MIC of SHHT was 100 µg/mL and the expression of cagA gene was decreased about 25 folds in the presence of SHHT. H. pylori urease was inhibited 90% by SHHT. SHHT inhibited H. pylori adhesion on AGS cell in a concentration dependent manner. mRNA expression of iNOS and IL-8 and the production of NO and IL-8 were significantly decreased in the presence of SHHT. In conclusion, SHHT showed anti-Helicobacter activity and has potent anti-inflammatory effect on H. pylori-induced inflammation in human gastric epithelial AGS cells.

The Association of Relative Risk of High Risk HPV with Cervical Abnormalities in Bahrain.

Background: Th e study was conducted to see the association of cervical abnormalities in relation to high risk Human Papilloma Virus (HR-HPV) infection at King Hamad University Hospital, Bahrain. Materials and Methods: It was a retrospective cohort study completed in 3 years at King Hamad University Hospital, Department Pathology Lab and other private Hospitals of Bahrain. Patients of Obstetrics and Gynecology out patient’s clinic and other requesting private hospitals were included in this study.A non-probability purposive sampling technique was used for this retrospective review of 160pathology reports and HPV cervista reports. Data was collected from I-Seha and patients Al-care, and was transferred and assessed SPSS-version 22. Results: Th ere were 160 cases in total, who were examined for HPV–HR DNA using Cervista molecular testing. Th ere were 73 cases were Positive for HPV and 87 cases negative for HPV. Th e minimum age of patiesnt's was 20 years while the max was 70 years. Th e mean age was 42.5 years. HR-HPV was detected in (100%) all cervical HSIL cases and in 71% of LSIL cases. Cervical intraepithelial lesion CIN2/3+ was signifi cantly associated with HR-HPV positive cases. Compared to HPV positive cases, here was no cervical intraepithelial lesion (CIN) of any grade found in HR_HPV negative cases. Th ere were only four cases with LSIL found to be R-HPV positive, which may be associated with Low-risk HPV infection. Conclusion: Th ere was strongest association of cervical neoplastic lesions with high risk HPV to control.

Apoptosis of human gastric carcinoma AGS cells induced by cecropinXJ / 中国药理学通报

Aim To investigate the apoptosis of human gastric carcinoma AGS cells induced by cecropinXJ. Methods Human gastric carcinoma AGS cells and human normal epithelial cells GES-1 were co-cultured with different concentrations of cecropinXJ ranging from 0. 01 to 1 000 mg·L-1 for 24 h. MTT assay was used to observe the effects of cecropinXJ on the proliferation of AGS cells and GES-1 cells. The ultrastructural changes of the AGS cells were observed by transmission electron microscopy. Hoechst staining was used to de-tect cell apoptosis. The changes of intracellular reac-tive oxygen species ( ROS) and mitochondrial potential were analysed by flow cytometery. The expression of Bax, Bcl-2, caspase-3 and cytochrome C in mRNA level was investigated by qRT-PCR. Western blot was used to determine the protein expression of Bax, Bcl-2, caspase-3 and cytochrome C. Results CecropinXJ significantly suppressed the proliferation of AGS cells in vitro (P<0. 05) in a dose-dependent manner, IC50 =61. 19 mg·L-1 , but had no inhibitive effects on the proliferation of GES-1 cells. After treatment for 24 h, cecropinXJ induced AGS cells nuclear condensation, and increased ROS production, disrupted mitochondri-al integrity. The results of qRT-PCR and Western blot demonstrated cecropinXJ could up-regulate the expres-sion of Bax and down-regulate the expression of Bcl-2 , promote the release of cytochrome C and activate caspase-3. Meanwhile, cecropinXJ promoted caspase-3 activity in a dose-dependent manner, and cell death ratio of AGS cells induced by cecropinXJ was signifi-cantly reduced by caspase-3 and caspase-9 specific in-hibitors treatment. Conclusion CecropinXJ can in-duce apoptosis of AGS cells by downregulating Bcl-2 , upregulating Bax and activating caspase-3 , which may be one of its anti-tumor mechanisms.

Effects of ω-3 polyunsaturated fatty acids on the proliferation of human gastric cancer cell line AGS / 西安交通大学学报(医学版)

ABSTRACT:Objective To investigate the effects of ω-3 polyunsaturated fatty acids on the proliferation of human gastric cancer cell line AGS and the possible mechanisms.Methods Human gastric cancer line AGS and human microvascular epithelial cell HMEC-1 were treated with different concentrations of docosdhexaenoic acid (DHA)and eicosapentaenoic acid (EPA).The inhibition of cell proliferation was evaluated by MTT assay and cell morphology.Flow cytometry was used to detect the cell cycle change.The expressions of mitochondrial respiratory membrane protein complex Ⅰ,Ⅱ and V were analyzed with Western blot.Results DHA and EPA could markedly inhibit the proliferation of AGS in significant time-dependent and concentration-dependent manners (P 0.05).Conclusion ω-3 PUFAs can selectively inhibit the growth and proliferation of human gastric cancer cell line AGS.These effects may be as-sociated with arresting cell cycle in G0/G1 phase and inhibiting the energy metabolism of AGS cells.

Production of specific IgY Helicobacter pylori recombinant OipA protein and assessment of its inhibitory effects towards attachment of H. pylori to AGS cell line

PURPOSE: The common triple therapy for Helicobacter pylori is challenged by the increasing cases of antibiotic resistant infections, raising the need to explore alternative therapies. Oral administration of egg yolk immunoglobulin Y (IgY) has been previously reported as a means of passive immunization therapy for H. pylori infections. In this work, we investigated the inhibitory effect of IgY on the attachment of H. pylori to AGS cell line. MATERIALS AND METHODS: Recombinant OipA was prepared. Hens were immunized with recombinant protein three times. IgY was purified from egg yolks of immunized hens using polyethylene glycol precipitation method. The inhibitory effect of the specific immunoglobulin was evaluated in AGS cell line infected with H. pylori. RESULTS: The presence of recombinant OipA (30 kD) was confirmed via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunization of hens was confirmed using enzyme-linked immunosorbent assay. The purified IgY from egg yolks were assessed using SDS-PAGE and confirmed by western blot. CONCLUSION: The results showed that IgY-OipA had inhibitory effect on attachment of H. pylori to AGS cell line and may be utilized as a therapeutic or prophylaxis material.

Effects of triptonoterpene methyl ether on proliferation and apoptosis of human gastric adenocarcinoma AGS cells / 中国药理学通报

Aim Toinvestigatetheeffectsoftriptonot-erpene methyl ether ( TME ) , a diterpene derived from the medicinal plant Triptergium wilfordii, on human gastric cancer AGS cell proliferation inhibition and ap-optosisinducedinvitro.Methods MTTassaywas used for screening tumor spectrum and detecting the vi-ability of AGS cells and normal human gastric epitheli-al cells GES-1 . Cell morphology was observed by light microscopy and AO / EB staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. JC-1 staining and fluorescence probe DCFH-DA were em-ployed to detect the changes of mitochondrial mem-brane potential and reactive oxygen species ( ROS ) . The effect of inhibiting AGS clonogenic survival was as-sayed by the method of plate clone formation. Western blot was used to analyse the expression of caspase-3 , caspase-8,Bcl-2andBax.Results MTTresults showed that TME exhibited significantly higher cytotox-icity to gastric cancer AGS cell line than to noncancer-ous cell line GES-1. IC50 for AGS of 48 h treatment was 23 . 85 μmol · L-1 . TME significantly inhibited colony formation and caused morphological changes in AGS cells. Annexin V-FITC / PI double staining showed the apoptotic rate increased. DCFH-DA stai-ning showed TME resulted in an increase in intracellu-lar ROS levels. Mitochondrial membrane potential de-creased after TME treatment. Western blot results showed that TME increased the proportion of Bax /Bcl-2 , with the activation of caspase-8 and caspase-3 . The broad-spectrum caspase inhibitor z-VAD-fmk pre-treatment reduced the expression of caspase-8 and caspase-3. TME enabled AGS cell cycle arrest in G0/G1phase.Conclusion TMEpossessespotenttumor selected toxicity and can induce apoptosis of AGS cells through cell cycle arrest, which is associated with Bcl-2 protein family.