1.T_1 and T_2 lymphocyte subset alterations in patients with colorectal cancer
Ming CUI ; Shan WANG ; Yingjiang YE ; Zhirong CUI ; Yang KE
Chinese Journal of General Surgery 2001;0(07):-
Objective The purpose of this study was to analyze the alterations of T_1、T_2 lymphocyte subsets in the peripheral blood of patients with colorectal cancer. MethodsTwenty patients with primary colorectal cancer were enrolled into this study, T_1 and T_2 in the peripheral blood were evaluated by detecting the intracellular interferon-? and interleukin-4 production with 4-color flow cytometry. ResultsThe percentage of T_1 and T_2 in the peripheral blood of cancer patients was lower significantly than healthy controls [(36?11)% and 3.3(1.9)% vs. (46?12)% and 4.1(3.1)%](P
2.The relationship between suppressor of zeste 12 expression and clinical features of gastric cancer
Hui ZHANG ; Yingjiang YE ; Zhirong CUI ; Shan WANG
Chinese Journal of General Surgery 2011;26(2):141-143
Objective To investigate the relationship between the expression of suppressor of zeste 12(SUZ12) and the clinicopathological parameters and prognosis in patients with gastric cancer. Methods SUZ12 protein expression levels in 97 cases of resected gastric cancer were detected by immunohitochemistry method, the relations between SUZ12 expression levels and the survival were estimated by Kaplan-Meier curve. Results The positive rate of SUZ12 expression in gastric cancer tissues was 43%, significantly higher than that (15%)in the adjacent noncancerous tissues( P = 0. 002). SUZ12-positive expression was significantly correlated with tumor differentiation ( P = 0. 018 ), lymph nodes metastasis ( P = 0. 023 ) and TNM staging(P = 0. 014). Gastric cancer patients with SUZ1 2-positive expression had worse prognosis than those with SUZ12-negative expression ( P = 0. 024). Conclusions SUZ12 is overexpressed in tissues of gastric carcinoma, SUZ12 is an independent prognosis factor of patients with gastric carcinoma.
3.Relationship between MiR-195 and DLL4 expression and clinical features of colorectal cancer
Hui ZHANG ; Chenggang WANG ; Zhirong CUI ; Yingjiang YE ; Jing ZHOU
Chinese Journal of General Surgery 2021;36(3):169-173
Objective:To observe the expression patterns of miR-195 and DLL4 in colorectal cancer, and to explore the relationship between miR-195 and DLL4 and clinicopathological parameters and prognosis of patients with colorectal cancer.Methods:The relative expression of miR-195 in 56 colorectal cancer tissues was detected by real-time fluorescent quantitative PCR the expression of DLL4 protein and tumor microvessel density (MVD) were detected by bloting and immunohistochemistry. Colon cancer cell line SW480 was treated with miR-195. The expression of DLL4, Jagged1, (the intracellular domain of notch, NICD), CyclinD1, Hes1, Bcl-2 and NF-kB were detected by bloting.Results:The expression level of DLL4 protein in colorectal cancer tissues was significantly higher than that in normal intestinal mucosa (30/56 vs.16/56, t=5.323, P=0.018). The expression level of miR-195 was significantly lower than that in normal intestinal mucosa (36/56 vs.20/56, t=2.371, P=0.008). The expression of DLL4 was negatively correlated with the expression of miR-195 ( r=- 0.881, P=0.015) , which was closely related to the differentiation degree, lymph node metastasis and TNM stage . The prognosis of patients with high expression of DLL4 was significantly worse than that with low expression of DLL4 ( P=0.013). The prognosis of patients with low expression of miR-195 was worse than that with high expression of miR-195 ( P=0.009) . Conclusion:The antagonistic expression of miR-195 and Notch might be closely related to the occurrence and development of colorectal cancer, which can be used as a new reference index for prognosis of colorectal cancer.
4.In vitro antitumor immune response induced by fusion of dendritic cells and metastatic colon cancer cells
Yu HE ; Shan WANG ; Yingjiang YE ; Feng XV ; Zhirong CUI
Chinese Journal of General Surgery 2001;0(08):-
Objective To detect the antitumor activity against SW620 and syngenic colon cancer SW480 by a fusion of human metastatic colon cancer SW620 cells and human peripheral blood-derived dendritic cells (DCs). Methods SW620 cells and human peripheral blood- derived DCs were fused with polyethylene glycol(PEG). The fusion cells were confirmed by "phenotypes determine, morphologic observation, and cytotoxic T lymphocyte response induced by the fusion were studied. Results Mature DCs with highly expressed surface markers (HLA-ABC, HLA-DR and CD80, CD86, CD83) were generated in vitro. The fusion of SW620 cells with DC resulted in a hybrid cell with morphologic as well as phenotypic characteristics of both DC and tumor cells. Flow cytometry showed that the highest fusion efficiency was 27. 12%. CTL assay demonstrated that the DC/SW620 fusion induced specific cytotoxic responses against the SW620 and SW480 cells. Conclusion The fusion of tumor cells with DCs induces tumor rejection.
5.Stat5b signaling pathway regulates the expression of Survivin and promotes apoptosis in human colon cancer cells
Xiangtao MA ; Liwei YU ; Shan WANG ; Ruyu DU ; Zhirong CUI
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: The purpose of the study was to examine colon cancer cell lines to determine whether Stat5b/Survivin plays an important role in the process of apoptosis in colon cancer cells. METHODS: Protein lysates were extracted from colon cancer cells. Human colon cancer cell line HT29 was transfected with Stat5b antisense oligonucleotide mediated by liposome. MTT assay was used to measure the proliferation. Flow cytometry was applied to analyze the cell cycle and apoptosis. EMSA was used to detect the activity of Stat5. Western blotting was applied to measure the expression of Stat5, p-Stat5, cyclin D1, Survivin, Bcl-2 and Bcl-xL. RESULTS: Targeting of Stat5 using antisense oligonucleotide against the translation site resulted in apoptosis and downregulaed the expressions of Stat5, p-Stat5, cyclin D1 and Survivin, but not Bcl-2 and Bcl-xL. CONCLUSION: Constitutive activation of Stat5 is associated with the carcinogenesis of colon cancer cells. Blocking of Stat5 signaling inhibits the expression of Survivin and induces apoptosis in colon cancer cells.
6.Janus kinase inhibitor AG490 inhibits gastric cancer cell invasiveness by down-regulation of MMPs and up-regulation of TIMPs
Bin LIANG ; Shan WANG ; Yingjiang YE ; Shen YANG ; Zhirong CUI
Chinese Journal of General Surgery 2001;0(10):-
Objective This study was to investigate the role of STAT3 signaling pathway in gastric cancer invasiveness. Methods Janus kinase 2 selective inhibitor AG490 was used to inhibit the activation of STAT3 signaling pathway. Matrigel-coated transwell was employed to estimate the invasiveness of BGG-823 cells. Alterations in the activity of STAT3 and expression of MMPs and TIMPs were measured by Western blot and RT-PCR. Results The activation of STAT3 signaling pathway was suppressed by AG490. AC490 significantly decreased the invasiveness of gastric cancer cell line BGC-823 through Matrigel-coated filters. Decreased invasion was associated with the inhibition of MMP-2, and MMP-9 at both mRNA and protein levels by RT-PCR and Western blot. Meanwhile, the expression of TIMP-1, and TIMP-2 were up-regulated at mRNA levels. Conclusion Inhibition of STAT3 signaling pathway in AG490 suppresses gastric cancer cell invasiveness through down-regulation of several invasion-related factors including MMP-2, and MMP-9 and up-regulation of their inhibitors.
7.Clinical significance of combined assay of serum tumor markers in patients with gastrocolonic carcinoma
Yingjiang YE ; Shan WANG ; Zhihai GAO ; Zhirong CUI
Chinese Journal of General Surgery 1997;0(04):-
Objective To evaluate the clinical significance of combined serum tumor markers assay in patients with gastric or colorectal carcinoma. Methods Serum level of 12 common tumor markers, including CA19-9,NSE,CEA,CA242 Jerritin,Beta-HCG,AFP,free-PSA,PSA,CA125,HGH and CA153, was measured with multi-tumor markers protein biochip detective system in 179 cases of gastric and colorectal carcinoma, 82 patients with benign digestive disease and 160 healthy volunteers. Results Cancer patients had significantly higher positive rates than that of two other controls (P
8.Effects of Dapper1 expression on surviving-mediated cell apoptosis in gastric carcinoma
Hui QIU ; Shan WANG ; Kewei JIANG ; Yingjiang YE ; Feng XU ; Zhirong CUI
Chinese Journal of General Surgery 2009;24(4):317-319
Objective To investigate the expressions of Dapper1 in gastric carcinoma and elucidate its relationship with survivin and its role in tumor cell apoptosis. Methods Dapper1 mRNA was detected with RT-PCR using specimens from 30 cases of gastric carcinoma and the corresponding normal gastric mucosa.The pcDNA3.1-Dpr1 plasmid was transfected into SGC-7901 cells with LipofectamineTM 2000.The effect of upregulation of Dpr1 on SGC7901 cell apoptosis was determined by flow cytometry.The downregulation of survivin、Dvl-2 and β-catenin protein expression were detected by Western blot analysis.Results Downregulation of Dpr1 gene expression was observed in 17(57%)of 30 human gastric cancer and the downregulation was significantly correlated with the depth of invasion and the degree of differentiation (P<0.05).Also,upregulation of Dpr1 mRNA and downregulation of survivin mRNA were detected after transfecting pcDNA3.1-Dpr1 plasmid in SGC7901 cells,which led to downregulation of survivin、Dvl-2、β-catenin protein and increase of the SGC7901 cell apoptosis rate from 2.89%to 13.96%.Conclusion Downregulation of Dp1l gene expression is common in human gastric carcinoma,and upregulation of Dpr1 results in significant inhibition of survivin expression which can induce apoptosis of SGC7901 cells.
9.TNF-α induces PIP3-mediated necroptosis in MLO-Y4 cells
Hongwang CUI ; Zhibin MENG ; Tao HUANG ; Kaizhong ZHU ; Zhirong ZHAO ; Yongjun ZHU
Chinese Journal of Pathophysiology 2017;33(8):1499-1505
AIM: To explore whether tumor necrosis factor-α (TNF-α) induces necroptosis in murine long bone osteocyte-like cell line MLO-Y4 and the possible mechanism.METHODS: The MLO-Y4 cells were divided into control group, TNF-α group, TNF-α+necrostatin-1 (Nec-1) group, TNF-α+Z-VAD group and TNF-α+receptor-interacting protein 3 (RIP3)-siRNA group.The death rate of MLO-Y4 cells was assessed by flow cytometry with Annexin V-FITC/PI staining.The morphological features of the cells were observed under transmission electron microscope (TEM).The protein levels of RIP1, RIP3 and cleaved caspase-3 were determined by Western blot.Finally, the numbers of total cells and RIP1-RIP3-positive cells were observed under laser scanning confocal microscope.The production of reactive oxygen species (ROS) in the cells was measured by DCFH-DA staining.RESULTS: Compared with control group, the apoptotic or necroptotic rate of the cells induced by TNF-α was increased significantly (P<0.01).The increased apoptotic or necroptotic rate was dramatically reduced by treating with Nec-1, Z-VAD or RIP3-siRNA transfection (P<0.01).In TNF-α group and TNF-α+Z-VAD group, a lot of MLO-Y4 cells with typical necroptotic morphological features were observed under TEM.However, obvious necroptotic cells were not found in Nec-1 or RIP3-siRNA treatment group.The protein level of RIP1 in the cells treated with Nec-1 was sharply lower than that in TNF-α group (P<0.01).However, Z-VAD did not reduce the elevated levels of RIP1 and RIP3.RIP3-siRNA effectively down-regulated the protein level of RIP3 compared with TNF-α group (P<0.01).Nec-1 effectively down-regulated the protein levels of RIP1 colocalized with RIP3 compared with TNF-α group (P<0.01).However, Z-VAD did not reduce the levels of RIP1 colocalized with RIP3.Nec-1, Z-VAD and RIP3 siRNA significantly decreased the ROS levels (P<0.01).CONCLUSION: TNF-α induces the necroptosis of MLO-Y4 cells.RIP3 play vital roles in the cell necroptotic signal pathway.ROS may be the executor of necroptosis of MLO-Y4 cells.
10.Molecular mechanisms involved in regulation of proliferation and apoptosis by Stat3 dominantnegative gene in colon cancer cells
Yong ZHAO ; Shan WANG ; Yingjiang YE ; Jing ZHOU ; Kewei JIANG ; Zhirong CUI
Chinese Journal of General Surgery 2001;0(09):-
Objective To study the influence of transferring a dominant-negative Stat3 gene, Stat3?on colon cancer cells' proliferation and apoptosis in vitro. Methods Cell culture of human colon cancer cell line SW480 and transient transfection were used to evaluate the effect of transferring Stat3?to cancer cells. Cell proliferation, cell cycle and apoptosis were quantified by MTT and flow cytometry, respectively. The mRNA expression of Stat3's target gene cyclin D1 and bcl-xL was detected by reverse transcription polymerase chain reaction. Independent t tests were used for data statistics. Results 36 h after Stat3?plasmids transfection, proliferation of SW480 cells was significantly inhibited (t =5. 216,P = 0.006); cell proportion of G0/G1 phase increased from 40.37% to 67.25% and early apoptosis cells increased from 5. 34% to 24. 42% ; mRNA expression of cyclin Dl and bcl-xL declined significantly (t = 5.288,P=0.010;t=3.517,P=0.025). Conclusion Blocking Stat3 signaling pathway by transfection of Stat3?plasmid inhibits the proliferation and promotes apoptosis of colon cancer cells, which provides a experimental foundation of Stat3 targeted colon cancer gene therapy.