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Objective To investigate the role of bone marrow mesenchymal stem cells (BM-MSCs) in the invasion and metastasis of gastric cancer cells and to explore its mechanism.Methods SGC7901 and KATO-Ⅲ gastric cancer cells were co-cultured with BM-MSCs respectively,and the invasion ability of SGC7901 and KATO-Ⅲ gastric cancer cells were detected by Transwell assay.Secondly,CD133 + and CD133-cells were sorted from KATO-Ⅲ gastric cancers and co-cultured with BM-MSCs respectively to compare their changes in invasiveness.Meanwhile,the expressions of p-AKT and epithelial-mesenchymal transition (EMT) relative factors in gastric cancer cells were detected by Western-blot.The role of CD133 in BM-MSCs affecting the ability of invasion of gastric cancer cells was further vertified by the overexpression of CD133 in SGC7901 cells.SPSS17.0 software was used for statistical processing,and the stand deviation of the measurement data were expressed as the standard deviation,independent sample t test was conducted.Results The invasiveness of co-cultured SGC7901 and KATO-Ⅲ cells was significantly enhanced.The invasive ability of KATO-Ⅲ CD133+ cells co-cultured with BM-MSCs tended to increase more significantly than that of co-cultured CD133 cells[(259.0 ± 24.0)vs (58.0 ±5.6),P < 0.001].The expressions of p-AKT,Snail and N-cadherin were significantly increased in co-cultured CD133+ cells (P =0.003,P =0.003,P =0.002),while the expression of E-cadherin was reduced (P =0.021).After co-cultured with BM-MSCs,the expression of E-cadherin was also reduced in CD133-cells (P =0.005),but the expressions of p-AKT,Snail and N-cadherin were no significantly changes (P =0.744,P =0.277,P =0.295).SGC7901 co-cultured with BM-MSC after overexpression of CD133 showed higher i nvasiveness than blank control group[(239.3 ± 24.0) vs (103.3 ± 15.5),P < 0.001].The expressions of p-AKT,Snail and N-cadherin were significantly increased when co-cultured with BM-MSCs in the group of CD133 overexpression (P =0.001,P =0.001,P =0.001),while the expression of E-cadherin was significantly decreased(P =0.003).The expressions of Snail and N-cadherin were also significantly increased after co-cultured with BM-MSCs in the blank control group (P =0.001,P =0.004),and the expression of E-cadherin was significantly decreased (P =0.018),while the expression of p-AKT was not significantly changed (P =0.193).Conclusions BM-MSCs can enhance the invasion and metastasis of gastric cancer cells by promoting the EMT of gastric cancer cells.CD133 may be involved in the regulation of EMT in gastric cancer cells through the PI3K/AKT signaling pathway.
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Objective To compare the curative effect of tissue-selecting therapy stapler and procedure for prolapse and hemorrhoids in the treatment of patients with stage Ⅲ to Ⅳ hemorrhoids.Methods The patients with stage Ⅲ to Ⅳ hemorrhoids who underwent prolapse and hemorrhoids or tissue-selecting therapy stapler surgery in the department of General Surgery,Shanghai Ninth People's Hospital and Xinhua Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Chongming Branch,from Jan.2013 to Jun.2014 were accepted and allocated to prolapse and hemorrhoids or tissue-selecting therapy stapler group.The peri-operative parameters about operative time,blood loss,postoperative hospital stay and the time required to return to normal activity were compared by t test,The postoperative complications including pain assessment and the incidence of postoperative bleeding,urine retention,faecal urgency,fecal incontinence,anal stenosis,rectovaginal fistula and recurrence rate were compared by t test and chi-square test.Rank sum test was used to compare the recurrence rate and patient's satisfaction between the two groups.Results The operation time,intraoperative bleeding volum,postoperative hospital stay and the time required to return to normal activity in the procedure for prolapse and hemorrhoids group were signifcantly higher than those in the tissue-selecting therapy stapler group (P =0.021,P =0.003,P =0.001,P <0.001).The pain score of procedure for prolapse and hemorrhoids group were all higher than those of the tissue-selecting therapy stapler group in the first post-operative defecation and in post-operative 24 hours and 72 hours (all P < 0.001).The incidence of faecal urgency of the procedure for prolapse and hemorrhoids group in post-operative 1 month (18.6%) was higher than that of the tissue-selecting therapy stapler group (6.6%) (P =0.036).There were no statistically significant differences in the incidence of postoperative bleeding,urinary retention,recurrence rate and patient's satisfaction between two group (P > 0.05).Conclusion Tissue-selecting therapy stapler was superior to the procedure for prolapse and hemorrhoids in operation time,intraoperative blood loss,postoperative pain and the incidence of faecal urgency.Long-term results demonstrate that tissue-selecting therapy stapler and prolapse and hemorrhoids have similar effectiveness.
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Objective To compare the curative effect of tissue-selecting therapy stapler and procedure for prolapse and hemorrhoids in the treatment of patients with stage Ⅲ to Ⅳ hemorrhoids.Methods The patients with stage Ⅲ to Ⅳ hemorrhoids who underwent prolapse and hemorrhoids or tissue-selecting therapy stapler surgery in the department of General Surgery,Shanghai Ninth People's Hospital and Xinhua Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Chongming Branch,from Jan.2013 to Jun.2014 were accepted and allocated to prolapse and hemorrhoids or tissue-selecting therapy stapler group.The peri-operative parameters about operative time,blood loss,postoperative hospital stay and the time required to return to normal activity were compared by t test,The postoperative complications including pain assessment and the incidence of postoperative bleeding,urine retention,faecal urgency,fecal incontinence,anal stenosis,rectovaginal fistula and recurrence rate were compared by t test and chi-square test.Rank sum test was used to compare the recurrence rate and patient's satisfaction between the two groups.Results The operation time,intraoperative bleeding volum,postoperative hospital stay and the time required to return to normal activity in the procedure for prolapse and hemorrhoids group were signifcantly higher than those in the tissue-selecting therapy stapler group (P =0.021,P =0.003,P =0.001,P <0.001).The pain score of procedure for prolapse and hemorrhoids group were all higher than those of the tissue-selecting therapy stapler group in the first post-operative defecation and in post-operative 24 hours and 72 hours (all P < 0.001).The incidence of faecal urgency of the procedure for prolapse and hemorrhoids group in post-operative 1 month (18.6%) was higher than that of the tissue-selecting therapy stapler group (6.6%) (P =0.036).There were no statistically significant differences in the incidence of postoperative bleeding,urinary retention,recurrence rate and patient's satisfaction between two group (P > 0.05).Conclusion Tissue-selecting therapy stapler was superior to the procedure for prolapse and hemorrhoids in operation time,intraoperative blood loss,postoperative pain and the incidence of faecal urgency.Long-term results demonstrate that tissue-selecting therapy stapler and prolapse and hemorrhoids have similar effectiveness.
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Tumor initiating cells are the source of the tumor,which have the characteristics of stem cells,such as self-renewal capacity,unlimited proliferation,multi-directional differentiation,DNA repair activity and resistance to apoptosis.The tumor initiating cells involved in chemotherapy drug resistance,traditional chemotherapy drugs is difficult to kill them,studies have shown that the tumor initiating cells involved in chemotherapy drug resistance.Tumor initiating cells are the significant factor of drug resistance and relapse in the tumor.tumor initiating cells resistance mechanism is complex,not only involved in tumor generally resistant characteristics,but kept natural resistance properties of stem cells.Tumor initiating cells resistant mechanism is controversial.This review introduces the resistance mechanisms of tumor-initiating cells both in tumor initiating cells-intrinsic and tumor initiating cells-extrinsic aspects based on the domestic and international relevant literatures.By revealing the drug resistance mechanism,it can be used to predict the effect of clinical chemotherapy,and now the article will review the research progress of the drug resistance mechanism of tumor initiating cells.
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Objective This study aimed at observing expression and clinical significance of metadherin in gastric adenocarcinoma and exploring the potentially regulating mechanism of metadherin in invation and migration of gastric cancer.Methods Expressions of metadherin and E-cadherin in primay lesion of gastric cancer were detected by immunohistochemistry and their correlation to clinicopathologic characteristics and prognosis were analyzed by Chi-square tests.Transwell assay and wound healing assay were applied for the ability of invasion and metastasis in gastric cancer cells.Then,the down-regulatied metadherin expression in MKN45 cells by RNA interference (siRNA) was carried out and furthermore,the regulation role of metadherin in epithelial-mesenchymal transition was analyzed also in invasion and migration of gastric cancer cells.Results The positive expression of metadherin was correlated to invading depth (P =0.029),lymph node metastasis (P =0.001),TNM stage (P =0.014) and inhibiting E-cadherin expression (P =0.001).The patients with positive metabherin shared poorer prognosis.Furthermore,the down-regulated metabherin in MKN45 cells would result in the increasing expression of E-cadherin,as well as decreasing expression of N-eadherin,Slug and Snail.At the same time,the abilities of invasion (P =0.027) and migration (P =0.008) of MKN45 cells was decreased.Conclusion metabherin induces EMT in metastasis of gastric adenocarcinoma via activating either Slug or Snail but not twist,which would result in the poorer prognosis.
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<p><b>OBJECTIVE</b>To investigate the regulatory mechanism of bone marrow mesenchymal stem cells(BMSC) on the biological profiles of KATO-III( cell lines of gastric cancer.</p><p><b>METHODS</b>Transwell cubicle was applied to build the co-cultured model in non-contact style. The differences of cell proliferation and the resistance of anti-tumour drug (5-fluoropyrimidinedione, 5-FU and Cisplatin, CDDP) between co-cultured group and single cultured group were evaluated by Cell Counting Kit 8-assay(CCK-8). The invasion ability was detected by Transwell assay. The expressions of stem cell makers, apoptosis-related factors and epithelium-mesenchymal transition (EMT)-related factors were detected by RT-PCR.</p><p><b>RESULTS</b>The proliferation ability of KATO-III( cells in co-cultured group was significantly stronger than that in single cultured group. The growth rate of KATO-III( cells in co-cultured group was significantly higher than that in single cultured group after treatment of 5-FU and CDDP(P<0.05). The mRNA expression level of Bcl-2 was significantly higher in co-cultured group KATO-III( cells(P<0.05), while the mRNA expression level of Bax was significantly lower in co-cultured group KATO-III( cells(P<0.05) in comparison with those in single cultured group. As compared to KATO-III( cells in single cultured group, the number of infiltrating-membrane cells was significantly higher (37.33±5.22 vs 14.56±2.54, P<0.01) in co-cultured group, and the mRNA expression levels of Snail and N-cadherin were significantly higher in co-cultured group KATO-III( cells (P<0.05), while the mRNA expression level of E-cadherin was significantly lower in co-cultured group KATO-III( cells (P<0.05). The expressions of CD133, Nanog and Sox-2 mRNA in co-cultured group KATO-III( cells were significantly higher than those in single cultured group(P<0.05).</p><p><b>CONCLUSIONS</b>In co-cultured model sharing non-contact style, BMSC can enhance such properties of KATO-III( gastric cancer cells as the proliferation, the invasion and the chemoresistance. Furthermore, the regulatory mechanisms may be related to the increase of the expressions of some stem cell markers in gastric cancer cells.</p>
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Humanos , Antineoplásicos , Apoptosis , Células de la Médula Ósea , Cadherinas , Línea Celular Tumoral , Proliferación Celular , Cisplatino , Técnicas de Cocultivo , Transición Epitelial-Mesenquimal , Fluorouracilo , ARN Mensajero , Células Madre , Neoplasias GástricasRESUMEN
The chemokine CXCL12 and its receptor CXCR4 played a very important role in the gastric carcinoma,which promoted the growth,invasion,and metastasis of the gastric carcinoma through various of signal pathways.In this paper,the literatures related CXCL12,signal pathways and gastric carcinoma were retrieved and explored.And the authors present the research progress concerning the roles of CXCL12 and its regulating mechanism in neoplasm,especially in gastric cancer.
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Objective: To investigate the influence of CD133+expression on patients' survival and resistance of CD133+cells to anti-tumor agents in gastric cancer (GC). Methods: Influence of CD133 expression on prognosis was analyzed employing sam-ples from patients with GC. GC cell lines were utilized to separate CD133+and CD133?subpopulations by immunomagnetic separation and to analyze the biological features of two subpopulations in vitro and in vivo, especially in resistant to anti-tumor reagents and its apoptotic mechanism. Results: The lower CD133+group showed a significantly better survival compared with the higher CD133+group. The highest content of CD133+subpopulations for KATO-III cells had stronger proliferative ability than CD133?subpopulations. A single CD133+cell was capable of generating new cell colony and the tumorigenicity rate in nude mice was 100% for CD133+ clonal spheres or for CD133+ cells, but 0% for CD133? cells. Furthermore, the higher expression levels of Oct-4, Sox-2, Musashi-1 and ABCG2 in CD133+ clonal spheres were identified compared with CD133+ cells or CD133? cells. Under the treatment of anti-tumor reagents, CD133+ cells had lower suppression rates compared with CD133? cells while lower level of Bcl-2 and higher level of Bax were found in CD133+cells compared with CD133?cells. Conclusions: The patients with lower CD133+expression had a better survival. Enriched CD133+ cells in clonal sphere shared the ability to be self-renewable, proliferative, tumorigenic and resistant to anti-tumor agents as probably regulated by Bcl-2 and Bax.
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<p><b>OBJECTIVE</b>To explore the relationship between CD133(+) subsets cells in human gastric cancer (GC) and molecules of drug resistance and their sensitivity to 5-FU.</p><p><b>METHODS</b>Three gastric cancer cell lines therein KATO-III(, SGC7901 and MKN45 were sorted by immunomagnetic beads cell sorting method. Then above cell lines were further divided into un-sorted GC cells, CD133(+) subgroup and CD133(-) subgroup. The expressions of CD133, P-gp, Bax and Bcl-2 were determined by RT-PCR, Western blot and immunoflurescence. Meanwhile, the sensitivity to 5-FU of three subgroups was detected by CCK-8 Kit. The apoptosis induced by 5-FU in three subgroups was determined by Hoechst 33258.</p><p><b>RESULTS</b>Expressions of CD133 in three CD133(+) subgroups were significantly higher than those in un-sorted GC cells and CD133(-) subgroup (all P<0.05). Expressions of P-gp and Bcl-2 in the three GC cell lines were different (all P<0.05). There were significant differences of expressions of P-gp, Bcl-2 and Bax among CD133(+) cells, un-sorted GC cells and CD133(-) cells (all P<0.05). CCK-8 detection showed that CD133(-) subgroup of MKN45 GC cell line was more sensitive than CD133(+) cells to 5-FU (P<0.05). Hoechst 33258 staining showed that there were more apoptotic cells in CD133(-) subgroup as compared to other two subgroups, and the least apoptotic cells were observed in CD133(+) subgroup of MKN45 GC cell line (P<0.05). CD133 sirna was transfected into MKN45 GC cell line and could down-regulate the expressions of CD133, P-gp, Bcl-2 and p-Akt, while the expression of Bax increased (all P<0.05).</p><p><b>CONCLUSIONS</b>CD133 may contribute to the resistance of GC cells to chemotherapy drug through P-gp, Bcl-2 and Bax. PI3K/Akt signal pathway may be involved in this process.</p>
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Humanos , Antígeno AC133 , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP , Antígenos CD , Metabolismo , Antineoplásicos , Farmacología , Apoptosis , Línea Celular Tumoral , Resistencia a Antineoplásicos , Fluorouracilo , Glicoproteínas , Metabolismo , Péptidos , Metabolismo , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , ARN Interferente Pequeño , Neoplasias Gástricas , Quimioterapia , Metabolismo , Patología , Proteína X Asociada a bcl-2RESUMEN
Long non-coding RNA (LncRNA) is a kind of length greater than 200 nucleotides not encode any proteins RNA molecules.Although not encode any proteins,LncRNA through chromatin modification,transcription activation and interference et al.involved in a variety of control process in the cell.In recent years,the study found that the expression of a variety of LncRNA abnormalities in gastric cancer.These changes are closely associated with the development and prognosis of gastric cancer,regulate the expression of related LncRNA can significantly improve the prognosis of patients.Targeted to regulate the expression of LncRNA can provide new strategies for treatment of gastric cancer.
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Nowadays,laparoscopy has already been widely applied to the diagnosis and treatment of kinds of diseases,improving the outcome of the patients with its less invasive advantages.In the treatment of gastric cancer,doctors always choose different methods of laparoscopic gastrectomy(LG) according to the tumor's location and its invasion and metastasis.But LG hasn't got widely application in the treatment of gastric cancer,which is meanly attributed to the exist of some Technical problems,failing to control the complications very well and the lack of evidence of long-term oncological outcomes.But with the development of new technology and the accumulation of the surgeons' experience,these problems might be conquered to a certain extent.In this review,the author will summary the application status of LG in the treatment of gastric cancer.
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Objective To investigate the clinicopathologic characters and prognostic value of epithelial mesenchymal transition (EMT) related proteins Snail and chemokine receptors 4(CXCR4) in gastric cancer.Methods The expressions of Snail and CXCR4 in the gastric tissues of 50 patients with gastric cancer were detected by Immunohistochemistry.The relation between the expressions of Snail and CXCR4 and the clinicopathologic characters were analyzed.The relative relationship between Snail expression and CXCR4 expression were identified by Spearman method.The correlation between the expressions of Snail and CXCR4 with the survival was analyzed by Kaplan-Meier method.Results The expression of Snail and CXCR4 expression were positive in 31 of 50 cases (62%) and 29 of 50 cases (58%) respectively.The expression levels of Snail and CXCR4 in the gastric cancer patients with diameter more than 5 cm,worse tissue differentiation,vascular invasion,lymphatic vessel invasion,lymph nodes metastasis,and T3 + T4 staging were significantly higher than those in the patients with diameter less than 5 cm,without vascular invasion,lymphatic vessel invasion,lymph nodes metastasis,and T1 + T2 staging (P < 0.05).The Expression of Snail and CXCR4 was positively Correlated(r =0.330,P =0.014).The co-expression of Snail and CXCR4 was significantly associated with poorer prognosis and was applied as an independent prognostic factor for gastric cancer (P =0.001).Conclusions The co-expression of Snail and CXCR4 was the independent prognostic factor for gastric cancer.The combining effect of EMT and CXCR4 may promote the progressions and metastasis of gastric cancer.Therefore,it may be of an effective way for the metastasis of gastric cancer to adjust these targets of the Snail and CXCR4.
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Cancer stem cells are special stem cells and are found recently in many tumors,and have the capability of self renewal and differentiation and can gradually differentiate into matured tumor cells to form the primary lesion and the various metastasis lesions of tumors.As well known,cancer stem cells shared strong ability of resistant to chemotherapy and/or radiotherapy,which is mainly related to many kinds of molecules and theirs relative regulation in niche regarding to cancer stem cells.Currently,the strategies of some novel therapies are conducted to detect the targeting sites with relevant to the microenvironments of cancer stem cells,such as the angiogenesis and epithelial-mesenchymal transition of cancer stem cells.Furthermore,insights into drug-resistance of cancer stem cells and its niche may offer new therapeutic strategies for the treatment of tumors.
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Objective To study the resistances of CDl33 + subset purified from gastric cancer cell line to chemotherapy drugs and the mechanism of this resistance regarding to the mRNA expressions of both Bcl-2 and BAX in relation to the relative apoptotic genes.Methods CD133 + subset and CD133-subset were purified from KATOⅢ cell linc by magnetic activated cell sorting.The proliferating ability of these two subsets resistantnt to 5-FU,Cisplatin(DDP,PDD) and Etoposide was checked and compared by CCK-8 test.The apoptotic changes of these two subsets regarding to the expression of mRNA of both Bcl-2 and BAX were also analized by RT-PCR.Results In CD133 + subset,the contant percentage of CD133 + expression rate was 90% via analysis of flow cytometye.Twelve hours after treatment of5-FU,DDP and VP-16,the cells in both CD133 + subgroup and CD133-subgroup would gradually start to change in apoptotic morphology.The growth inhibiting rate by CCK-8 measurement for 5-FU,DDP and VP-16 groups in CD133 + subgroup was significantly lower than that in CD133-subgroup.The data under different treatment respectively was,5-FU:(30.56 ± 1.99) %-(88.60 ± 1.95) % vs (32.81 ± 2.67) %-(95.73±2.12)%,P=0.045,cisplatin:(45.89 ±3.64)%-(81.20 ± 1.18)% vs (50.21 ±3.22)%-(90.46±1.89)%,P=0.043,VP-16:(37.21 ±3.80)%-(78.49 ±3.22)% vs (35.55 ±3.23)%-(89.32 ±-3.54) %,P =0.048).After treatment of these three kind of anti-tumour drugs,the expression level of Bcl-2 mR-NA decreased significantly and the expression level of BAX mRNA increased significantly in both CD133 + subgroup and CD133-subgroup.However,these changing ranges of Bcl-2 mRNA and BAX mRNA were more obvious in CD133 + subgroup in comparison with those in CD133-subgroup.Conclusions In some degree,resistent potentiality of CD133 + cells to 5-FU,DDP and VP-16 has been identified,which may probably be due to the up-regulation of the expression of BAX and down-regulation of the expression of Bcl-2.
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Objective To analyze the clinical and pathological characteristics and its surgical management strategy for colorectal high-grade intraepithelial neoplasia (HGIN).Methods Eighty-two cases with colorectal tumors diagnosed as colorectal HGIN based on colonoscopic biopsy between January 2005 and December 2012 were enrolled in the study.The clinicopathological data of all the patients was collected and analyzed.Of the 82 cases,71 cases had radical colorectal surgery,1 cases had Miles operation after previous transanal excisions,3 had transanal local excisions,8 cases had palliative surgery.The surgical specimens were all examined pathologically and compared with the preoperative diagnosis of colonoscopic biopsy of all the patients.Results Three cases (3.7%) were pathologically diagnosed as high-grade intraepithelial neoplasia,their average diameter was 1.5 cm.The other 79 (96.3%) cases were diagnosed as adenocarcinoma,with an average diameter of 4.7 cm.The difference in tumor size was statistically significant (P < 0.01).Comparison of pre-and post-operative specimens showed poor consistency,the Kappa value was O.104.Significant analysis showed a correlation between cancerous change to tumor size and depth of invasion.In the 79 cases confirmed as adenocarcinoma,liver metastasis occurred in 8 cases,regional lymph nodes metastasis in 31 cases (39.2%).Of the 33 cases with rectal tumors,30 cases (90.9%) were pathologically diagnosed as adenocarcinoma after operation.Conclusions Much attention should be payed to the pathological diagnosis in colorectal intraepithelial neoplasia,especially in the HGIN.We have found that of the cases first diagnosed as HGIN,approximately 96.3% already have invasion adenocarcinoma.Most cases had reginonal lymph nodes metastasis.Liver metastasis had been occurred.thus active surgical measures should be taken.If the location of the tumor was not involved to anal sphincter,or cases with tumors larger than 3 cm was diagnosed,in highly suspected cases with malignant potiential,radical surgery is recommended.For tumors located at the lower rectum,the final decision should be made only after repeated endoscopic or transanal biopsy.
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Objective To investigate if TGF-β1 induces epithelial-mesenchymal transition (EMT) and promotes the obtaining of stemness characteristics in gastric cancer cell lines.Methods After KATO-Ⅲ cells were cultured with or without 5 ng/mL TGF-β1,the morphological change was observed and compared under phase-contrast microscopy.At the same time,the effect of TGF-β1 on the proliferation of KATO-Ⅲ cells was detected by CCK-8.On the other hand,the mRNA and protein' s expressions of EMT-related factors,ESC markers and TICs markers were analyzed by RT-PCR and Western blotting methods too.Results TGF-β1 induced morphological alterations from epithelial to mesenchymal cells.The proliferation of KATO-Ⅲ cells was inhibited after treated with TGF-β1 (P < 0.05).After treated with TGF-β1,the relative mRNA expression levels of Snail (0.5219 ±0.0147) and N-cadherin(0.6640 ±0.0124) were higher than that in control group(0.2049 ±0.0214,P =0.004,0.2722 ± 0.0098,P =0.001),the relative protein expression levels of Snail (0.4769 ± 0.0234) and N-cadherin (0.5014 ± 0.0216) were higher than that in control group (0.2534 ± 0.0345,P =0.02,0.2026 ± 0.0268,P =0.009),while the relative E-cadherin mRNA and protein levels in TGF-β1 treated group (0.4701 ± 0.0215,0.1349 ± 0.0258) were lower than that in control group (0.6792 ± 0.0157,P =0.01 ; 0.6055 ± 0.0227,P =0.004),while the relative mRNA expressions of ESC markers such as Sox2,OCT4,Nanog in TGF-β1 treated group (0.594 ± 0.039、0.438 ± 0.033、0.489 ± 0.037) were higher than that in control group (0.143 ± 0.013,P =0.001,0.156 ± 0.025,P =0.001,0.325 ± 0.046,P =0.03),the relative mRNA expression levels of CD44 (0.437 ±0.037) and CD133(0.543 ±0.028) were higher than that in control group (0.247 ±0.024,P =0.000,0.139 ± 0.016,P =0.000),the relative protein expression levels of CD44 (0.429 ± 0.034) and CD133 (0.316 ±0.027) in TGF-β1 treated group were higher than that in control group (0.152 ± 0.014,P =0.000,0.110 ±0.010,P =0.000),cloning sphere-forming capacity was greatly enhanced after treated with TGF-β1 (P < 0.01).Conclusion TGF-β1 can induce EMT in KATO-Ⅲ cells and promote the obtaining of stemness characteristics in gastric cancer cell lines.
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Objective To compare the inhibition effects of three synthesized fragments used in small interfering RNA(siRNA) against CD133 gene in KATO-Ⅲ gastric cancer cells,and to study effects of suppressed CD133 on the proliferating ability of intervened cells.Methods Three fragments of siRNA were designed and synthesized targeted at the mRNA of CD133.Cell fluorescence counting under confocal laser scanning microscope was used to determine the transfection efficiency after transfection with the CD133FITC-siRNA.The knock-down effect of the CD133 gene was detected by RT-PCR and Western blotting.CCK-8 (cell counting kit-8 assay) was performed to measure the variation of the cell proliferative viability after the above-mentioned treatment.Results The transfection efficiency of siRNA was (85 ± 8) % in KATO Ⅲ Gastric cacer cell.All these three fragments of CD133 siRNA effectively inhibited the expression of CD133 gene,the inhibition rate being (11 ± 2) %,(19 ± 2) %,(24 ± 3) %respectively.Compared with the control group,the cell proliferation viability was restrained (42 ± 4)% in CD133siRNA-3 group (P <0.05).Conclusions CD133siRNAs were successfully transfected into KATO Ⅲ Gastric cacer cells and repressed the expression of CD133.Meanwhile,the CD133siRNA fragment 3 was screened from three CD133 siRNA,which has the best inhibition effect.The results provide preliminary evidence for the intereference of CD133+ gastric cancer cells subsequently.
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Objective To investigate the biological effect of epithelial-to-mesenchymal transition (EMT) under the treatment of transforming growth factor (TGF)-β1 on the human gastric cancer cell line SGC7901 in vitro,and to observe whether the TGF-β1 can generate the tumor initiating cells ability in SGC7901 or not.Methods SGC7901 cells were cultured with TGF-β1.The morphological change was observed.The effect on proliferation of SGC7901 cells was detected by CCK-8.The invasion assay was used to investigate the motility and the invasion ability of SGC7901 cells.Immuofluorescence was used to detect the expression of E-cadherin and N-cadherin.The mRNA and protein's expression levels of EMT-related factors and CD44 were analyzed by RT-PCR and Western blotting respectively.Results TGF-β1 induced morphological alterations from epithelial to mesenchymal cells.The proliferation of SGC7901 cells was inhibited,and the ability of motility and invasion of SGC7901 cells were greatly enhanced after being treated with TGF-β1.RT-PCR and Western blotting showed that the expression of Snail (P < 0.05),N-cadherin (P < 0.05) and CD44 (P < 0.05) were significantly increased while the expression of E-cadherin was decreased (P < 0.05).Conclusions TGF-β1 can generate the EMT.The CD44 expression was up-regulated.TGF-β1 can inhibit the proliferation and promote the motility and invasion ability of SGC7901 cells.
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The morbidity and mortality of gastric cancer stand in the first place among all kinds of malignant tumours in our country,the early diagnosis rate is still very low and the prognosis is unsatisfactory also.The expression of CD133 probably as a tumour stem cell marker is significantly higher in gastric cancer primary lesion than that in normal tissue and its higher expression corelates with the larger size of tumor,lymph node metastasis occurences and poorer prognosis.Meanwhile,the positive expression of CD133 may have some relationship with the angiogenesis,severer infiltration depth,worse differentiation degree and later TNM stage of gastric cancer.This paper aims at further recongnizing the specific expression of CD133 in gastric cancer primary lesion,both the relationship with initiating and clinical pathological features of gastric cancer,so as to bring a new research direction for early diagnosis and targeted therapy of stomach cancer.
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Gallbladder carcinoma is the most common malignant tumor in biliary system,the early diagnosis rate is still very low and the prognosis is extremely poor.At present,the pathogenesis of gallbladder is not clear.Recently,with the development of the neoplastic stem cell research on gallbladder carcinoma,the latest study showed SP positive cells could be separated from the GBC-SD cell.Further more,related researches have shown that gallbladder respectively with CD133+ cells,CD44+ cells,CD133+ CD44+ cells had possessed the cloned formation ability,the stronger growth ability,the differentiation capacity and forming-tumor ability.In this review,we will discuss the research of the relationship between the gallbladder carcinoma and neoplastic stem cells.