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.

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.

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.

Objective: To investigate the curative effect of low-frequency therapeutic instrument of hepatopathy combined with Reduning injection in clinical treatment for patients with scarlet fever and hepatitis injury. Methods: 94 patients with scarlet fever and hepatitis injury were divided into observation group (47 cases) and control group (47cases). Patients of control group received the therapy of Reduning injection combined with tablet of adefovir dipivoxil, while the patients of observation group received the therapy of therapeutic instrument of hepatopathy combined with the therapy of control group. The changes of curative effect, indicator of hepatic fibrosis and response rate between two groups were observed. Results: The total effective rate of the observation group (95.7%) was significantly higher than that of control group (70.2%) (x2=7.283, P<0.05). In observation group, the hepatic fibrosis hyaluronic acid, III type procollagen, IV type collagen and laminin of post-treatment were significant improvement than that of pre-treatment (t=3.42, t=2.83, t=2.74, t=2.52, P<0.05). While in control groups, only the laminin of post-treatment was significant improvement than that of pre-treatment (t=2.15, P<0.05). After treatment, CR, VR, BR and HBeAg of observation group were significant higher than that of control group (x2=5.235, x2=5.623, x2=4.993, x2=6.823, P<0.05). Conclusion: The combination of low-frequency treatment of hepatopathy and Reduning injuection for the treatment of patients with scarlet fever and hepatic injury has significant effect, and it can effectively enhance the conversion rate of HBeAg, and improve liver function of patients. Therefore, it is worthy to be further promoteed in the clinical application.

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.

OBJECTIVETo investigate the regulatory mechanism of bone marrow mesenchymal stem cells(BMSC) on the biological profiles of KATO-III( cell lines of gastric cancer.

METHODSTranswell 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.

RESULTSThe 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).

CONCLUSIONSIn 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.

Antineoplastic Agents ; Apoptosis ; Bone Marrow Cells ; Cadherins ; Cell Line, Tumor ; Cell Proliferation ; Cisplatin ; Coculture Techniques ; Epithelial-Mesenchymal Transition ; Fluorouracil ; Humans ; RNA, Messenger ; Stem Cells ; Stomach Neoplasms

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.

Objective To investigate the regulatory mechanism of bone marrow mesenchymal stem cells(BMSC) on the biological profiles of KATO-Ⅲcell lines of gastric cancer. Methods 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. Results 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). Conclusions 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.

Objective To investigate the regulatory mechanism of bone marrow mesenchymal stem cells(BMSC) on the biological profiles of KATO-Ⅲcell lines of gastric cancer. Methods 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. Results 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). Conclusions 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.

OBJECTIVETo explore the relationship between CD133(+) subsets cells in human gastric cancer (GC) and molecules of drug resistance and their sensitivity to 5-FU.

METHODSThree 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.

RESULTSExpressions 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).

CONCLUSIONSCD133 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.

AC133 Antigen ; ATP-Binding Cassette, Sub-Family B, Member 1 ; Antigens, CD ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Fluorouracil ; Glycoproteins ; metabolism ; Humans ; Peptides ; metabolism ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; RNA, Small Interfering ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; bcl-2-Associated X Protein