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Objective To investigate the relationship between X-ray injury cross-complementing protein 1 (XRCC1) gene polymorphism and prognosis in patients with triple-negative breast cancer (TNBC). Methods Patients with primary triple-negative breast cancer (TNBC) diagnosed in the Ninth People′s Hospital, Shanghai Jiaotong University School of Medicine from January 2013 to January 2015 were selected. Patients were selected for genotyping (XRCC1 gene Arg280His, Arg399Gln and Arg194Trp) and divided into survival group and death group according to the prognosis of patients. Logistic regression was used to analyze the relationship between XRCC1 genotype and prognosis. Results A total of 130 patients were enrolled in the study, with an average age of (50.4 ± 6.3) years. The mean follow-up time was (45 ± 13) months, including 62 patients with breast cancer-related deaths and 68 patients with survival. The patients in death group was older than those in the survival group [(52.6 ± 6.7) years vs. (48.3 ± 5.2) years, P < 0.01), and and the lymph node metastasis rate was higher [88.7%(55/62) vs.73.5%(50/68), P＝0.028]. The frequency of XRCC1 gene Arg399Gln genotype in the survival and death group was GG: 61.8% vs. 38.7%; GA: 32.4% vs. 41.5%; AA: 5.9% vs. 19.4%, P＝0.011.There were also statistical differences between the two groups in the frequency of allele, and the frequency of A allele was significantly higher in the death group than in the survival group (40.4% vs. 22.0%, P<0.01). For the additive model of the Arg399Gln polymorphism A allele, for each additional copy of the A allele, the risk was 1.443 times that before the increase (95% CI 1.174-1.793, P<0.01). After adjusting for age and lymph node metastasis, the A allele still significantly increased the risk of death ( OR＝1.533, 95% CI 1.254-1.903, P < 0.01). Conclusions The XRCC1 gene Arg399Gln polymorphism is associated with the prognosis of TNBC, and patients with the A allele have a poor prognosis.
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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 investigate the incidence of adult epilepsy patients with attention-deficit hyperactivity disorder (ADHD) and the characteristics of ADHD subtypes, and to explore the correlations between several epilepsy-related factors and ADHD. Methods According to corresponding inclusion criteria and exclusion criteria, 82 adult epilepsy patients from March 2012 to March 2016 were enrolled as study group. Eighty-two healthy adults were enrolled as the control group. Using the Chi-square test and Logistic regression analysis and the other statistical methods, the phenomenon of ADHD in adults with epilepsy and the factors that may contribute to the phenomenon was analyzed. Results The detection rate of ADHD in 82 adult patients with epilepsy was 26.8%(22/82), significantly higher than that in the heathly control group which was 4.9%(4/82),χ2=18.672, P<0.05. Symptomatic epilepsy 8/17 and the electroencephalogram showed multifocal discharge 6/14 had higher detection rate of comorbid ADHD and the difference was statistically significant (P<0.01). The results of binary Logistic regression analysis showed that epilepsy age of onset was possibly an important factor for epilepsy with ADHD (B=-0.891, P<0.05). Conclusions The risk of ADHD in adults with epilepsy is higher than that in the healthy adults and attention-deficit is the common subtype. The etiology, age of onset and electroencephalogram features of adults with epilepsy may be associated with the occurrence of ADHD.
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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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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 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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Humans , Antineoplastic Agents , Apoptosis , Bone Marrow Cells , Cadherins , Cell Line, Tumor , Cell Proliferation , Cisplatin , Coculture Techniques , Epithelial-Mesenchymal Transition , Fluorouracil , RNA, Messenger , Stem Cells , Stomach NeoplasmsABSTRACT
<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>
Subject(s)
Humans , AC133 Antigen , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Antigens, CD , Metabolism , Antineoplastic Agents , Pharmacology , Apoptosis , Cell Line, Tumor , Drug Resistance, Neoplasm , Fluorouracil , Glycoproteins , Metabolism , Peptides , Metabolism , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt , RNA, Small Interfering , Stomach Neoplasms , Drug Therapy , Metabolism , Pathology , bcl-2-Associated X ProteinABSTRACT
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 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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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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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.