Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunc...Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells(PBMCs)were isolated,then rhHMGB1 was added to PBMCs.Four-color flow cytometric(FCM)analysis was used for the measurement of intracellular cytokine including interleukin IL-4 and interferon IFN-?ELISA kits were employed for the determination of IL-2 and sIL-2R protein levels in cell culture supernatants.Results(1)Different stimulating time and dosage of rhHMGB1 did not alter the number of IFN-αpositive cells(Th1).rhHMGB1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th1 to Th2.(2)Compared with the untreated cells,when the cells were coincubated with rhHMGB1(10-100ng/ml)for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGB1.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.展开更多
AIM:To investigate the effect of high mobility group A2(HMGA2) gene silencing on gastric cancer MKN-45 cells in vitro.METHODS:HMGA2 short hairpin RNA(shRNA) expression plasmids were constructed,including a pair of ran...AIM:To investigate the effect of high mobility group A2(HMGA2) gene silencing on gastric cancer MKN-45 cells in vitro.METHODS:HMGA2 short hairpin RNA(shRNA) expression plasmids were constructed,including a pair of random scrambled sequences.Human gastric cancer cell line MKN-45 cells were divided into three groups:blank control group(non-transfected cells),transfected group(cells transfected with HMGA2 shRNA recombinant plasmid) and scrambled sequence group(transfected with random scrambled plasmid).Cells were transfected with HMGA2 shRNA recombinant plasmids and scrambled plasmid in vitro,and the cells transfection efficiency was assayed by fluorescence microscopy.The HMGA2 messenger RNA(mRNA) expression was detected by reverse transcription polymerase chain reaction,gastric cancer cells apoptosis was detected by flow cytometry,cell proliferation was detected by methyl thiazol tetrazolium,and the protein expression of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),P27,caspase-9 and B-cell leukemia/lymphoma-2(Bcl-2) were analyzed by Western blotting.RESULTS:Compared with the blank control group and the scrambled sequence group,the levels of HMGA2 mRNA and protein expression in the transfected group were significantly reduced(P < 0.05).The relative HMGA2 mRNA expression levels of the blank control group,transfected group and scrambled sequence group were 0.674 ± 0.129,0.374 ± 0.048 and 0.689 ± 0.124,respectively.The relative HMGA2 protein expression levels of the blank control group,transfected group and scrambled sequence group were 0.554 ± 0.082,0.113 ± 0.032 and 0.484 ± 0.123,respectively.Moreover,transfection with the scrambled sequence had no effect on the expression of HMGA2.After being transfected with shRNA for 24,48 and 72 h,the cell apoptotic rates of the transfected group were 21.65% ± 0.28%,39.98% ± 1.82% and 24.51% ± 0.93%,respectively,which significantly higher than those of blank control group(4.72% ± 1.34%,5.83% ± 0.13% and 5.22% ± 1.07%) and scrambled sequence group(4.28% ± 1.33%,7.87% ± 1.43% and 6.71% ± 0.92%).After 24,48 and 72 h,the cell proliferation inhibition rates in the transfected group were 31.57% ± 1.17%,39.45% ± 2.07% and 37.56% ± 2.32%,respectively;the most obvious cell proliferation inhibition appeared at 48 h after transfection.Compared with the blank control group and scrambled sequence group,after transfection of shRNA for 72 h,the protein expression levels of PI3K(0.042 ± 0.005 vs 0.069 ± 0.003,0.067 ± 0.05),Akt(0.248 ± 0.004 vs 0.489 ± 0.006,0.496 ± 0.104) and Bcl-2(0.295 ± 0.084 vs 0.592 ± 0.072,0.594 ± 0.109) were significantly reduced.The protein expression levels of P27(0.151 ± 0.010 vs 0.068± 0.014,0.060 ± 0.013) and caspase-9(0.136 ± 0.042 vs 0.075 ± 0.010,0.073 ± 0.072) were significantly upregulated.CONCLUSION:HMGA2 shRNA gene silencing induces apoptosis and suppresses proliferation of MKN-45 cells.展开更多
Objective:To explore the relationship of plasma homocysteine(Hcy),soluble intercellular adhesion molecule-1(sICAM-1)and high mobility group box 1 protein(HMGB1)with carotid intima-media thickness(c-IMT)in elderly pati...Objective:To explore the relationship of plasma homocysteine(Hcy),soluble intercellular adhesion molecule-1(sICAM-1)and high mobility group box 1 protein(HMGB1)with carotid intima-media thickness(c-IMT)in elderly patients with type 2 diabetes mellitus.Methods:A total of 100 elderly patients who were diagnosed as type 2 diabetes mellitus in Baogang Hospital of Inner Mongolia from June 2017 to May 2020 were chosen as research objects.According to c-IMT,they were divided into the normal group(n=35),the mild to moderate group(n=41)and the severe group(n=24).The expression levels of plasma Hcy,sICAM-1 and HMGB1 were compared between groups respectively.Pearson’s correlation coefficient was used to analyze the relationship of plasma Hcy,sICAM-1,HMGB1 with c-IMT.Results:The comparison in plasma Hcy,sICAM-1,HMGB1 and c-IMT among the three groups of patients was of statistical significance(p<.05).The results of correlation analysis showed that the expression levels of plasma Hcy,sICAM-1 and HMGB1 were positively correlated with c-IMT in elderly patients with type 2 diabetes mellitus(r=.627,.598,.614;p<.05).Conclusions:The expression levels of plasma Hcy,sICAM-1 and HMGB1 are abnormally increased in elderly patients with type 2 diabetes mellitus,and related to c-IMT,which can provide a strong evidence for clinical diagnosis and treatment by detecting their levels in clinical practice.展开更多
目的探讨High mobility group A2protein(HMGA2)的表达对非小细胞肺癌生长、转移的影响,与临床病理参数和细胞增殖的关系。方法应用免疫组化法检测38例非小细胞肺癌组织(23例鳞癌,15例腺癌)及癌旁的正常肺组织标本中HMGA2和Ki-67的表达...目的探讨High mobility group A2protein(HMGA2)的表达对非小细胞肺癌生长、转移的影响,与临床病理参数和细胞增殖的关系。方法应用免疫组化法检测38例非小细胞肺癌组织(23例鳞癌,15例腺癌)及癌旁的正常肺组织标本中HMGA2和Ki-67的表达。结果HMGA2和Ki-67在癌旁的正常肺组织中均不表达,而在肺癌组织中的表达阳性率分别为39.47%、44.74%,二者在肺癌组和癌旁的正常肺组织组之间差异有显著性(P<0.05)。HMGA2的表达在伴淋巴结转移的肺癌组织明显高于不伴有淋巴结转移的肺癌组织(P<0.05),而与其他临床病理参数包括肿瘤的分化程度、肿瘤大小和TNM分期以及细胞增殖指标Ki-67之间没有相关性。结论本研究显示HMGA2在非小细胞肺癌组织中异常表达,可能与肺癌的发生和进展有关。展开更多
Objective:To analyze effects of high mobility group AT-hook 2(HMGA2)on malignant degree,invasion,metastasis,proliferation and cellular morphology of ovarian cancer cells.Methods:Three methods were applied to observe t...Objective:To analyze effects of high mobility group AT-hook 2(HMGA2)on malignant degree,invasion,metastasis,proliferation and cellular morphology of ovarian cancer cells.Methods:Three methods were applied to observe the effect on HMGA2 expression in ovarian cancer cells and ovarian epithelial cells.Results:After the application of siRNA-HMGA2,number of T29A2-cell clones was decreased,there was significant difference compared with the negative control Block-iT.After application of let-7c,number of T29A2+cell clones was decreased significantly,however,after the application of Anti-let-7,the number of clones restored,and there was no significant difference compared with the negative control group.After interference,the number of T29A2-cells which passed through Matrigel polycarbonate membrane were significantly lower than the negative control group.After the treatment of siRNA-HMGA2,let-7c and sh-HMGA2respectively,growth and proliferation of T29A2-,T29A2+and SKOV3 were slower,and the phenomenon was most obvious in SKOV3.Stable interference of HMGA2 induced mesenchymalepithelial changes in the morphology of SKOV3-sh-HMGA2.Conclusions:HMGA2 can promote malignant transformation of ovarian cancer cells,enhance cell invasion and metastasis,and promote cell growth and proliferation of ovarian cancer cells,which can cause ovarian cancer to progress rapidly and affect the quality of life.展开更多
High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression oc...High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression occurrs in many kinds of cancers including colorectal cancer,suggesting that HMGA2 might play a critical role in the progression of various tumors.However,the exact role of HMGA2 in colorectal cancer has not been determined.To verify the essential role of HMGA2 in the growth and invasiveness of colorectal cancer,HMGA2 expression was down-regulated by RNA interference（RNAi） in SW480 cells.We observed that the knockdown of HMGA2 led to the significant inhibition of proliferation and invasion of SW480 cells in vitro.These results suggest that HMGA2 might play a crucial role in the progression of colorectal cancer,and be a potential therapeutic target for human colorectal cancer.展开更多
基金This study was supported by the National Natural Science Foundation of China(No.30672178)National Basic Research Program of China(No.2005CB522602)the National Natural Science Outstanding Youth Foundation of China(No.30125020).
文摘Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells(PBMCs)were isolated,then rhHMGB1 was added to PBMCs.Four-color flow cytometric(FCM)analysis was used for the measurement of intracellular cytokine including interleukin IL-4 and interferon IFN-?ELISA kits were employed for the determination of IL-2 and sIL-2R protein levels in cell culture supernatants.Results(1)Different stimulating time and dosage of rhHMGB1 did not alter the number of IFN-αpositive cells(Th1).rhHMGB1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th1 to Th2.(2)Compared with the untreated cells,when the cells were coincubated with rhHMGB1(10-100ng/ml)for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGB1.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.
基金Supported by The Natural Science Foundation of Guangxi,No. 2010GXNSFA013166the Key Project of Health Department of Guangxi,No.Zhong 2010021
文摘AIM:To investigate the effect of high mobility group A2(HMGA2) gene silencing on gastric cancer MKN-45 cells in vitro.METHODS:HMGA2 short hairpin RNA(shRNA) expression plasmids were constructed,including a pair of random scrambled sequences.Human gastric cancer cell line MKN-45 cells were divided into three groups:blank control group(non-transfected cells),transfected group(cells transfected with HMGA2 shRNA recombinant plasmid) and scrambled sequence group(transfected with random scrambled plasmid).Cells were transfected with HMGA2 shRNA recombinant plasmids and scrambled plasmid in vitro,and the cells transfection efficiency was assayed by fluorescence microscopy.The HMGA2 messenger RNA(mRNA) expression was detected by reverse transcription polymerase chain reaction,gastric cancer cells apoptosis was detected by flow cytometry,cell proliferation was detected by methyl thiazol tetrazolium,and the protein expression of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),P27,caspase-9 and B-cell leukemia/lymphoma-2(Bcl-2) were analyzed by Western blotting.RESULTS:Compared with the blank control group and the scrambled sequence group,the levels of HMGA2 mRNA and protein expression in the transfected group were significantly reduced(P < 0.05).The relative HMGA2 mRNA expression levels of the blank control group,transfected group and scrambled sequence group were 0.674 ± 0.129,0.374 ± 0.048 and 0.689 ± 0.124,respectively.The relative HMGA2 protein expression levels of the blank control group,transfected group and scrambled sequence group were 0.554 ± 0.082,0.113 ± 0.032 and 0.484 ± 0.123,respectively.Moreover,transfection with the scrambled sequence had no effect on the expression of HMGA2.After being transfected with shRNA for 24,48 and 72 h,the cell apoptotic rates of the transfected group were 21.65% ± 0.28%,39.98% ± 1.82% and 24.51% ± 0.93%,respectively,which significantly higher than those of blank control group(4.72% ± 1.34%,5.83% ± 0.13% and 5.22% ± 1.07%) and scrambled sequence group(4.28% ± 1.33%,7.87% ± 1.43% and 6.71% ± 0.92%).After 24,48 and 72 h,the cell proliferation inhibition rates in the transfected group were 31.57% ± 1.17%,39.45% ± 2.07% and 37.56% ± 2.32%,respectively;the most obvious cell proliferation inhibition appeared at 48 h after transfection.Compared with the blank control group and scrambled sequence group,after transfection of shRNA for 72 h,the protein expression levels of PI3K(0.042 ± 0.005 vs 0.069 ± 0.003,0.067 ± 0.05),Akt(0.248 ± 0.004 vs 0.489 ± 0.006,0.496 ± 0.104) and Bcl-2(0.295 ± 0.084 vs 0.592 ± 0.072,0.594 ± 0.109) were significantly reduced.The protein expression levels of P27(0.151 ± 0.010 vs 0.068± 0.014,0.060 ± 0.013) and caspase-9(0.136 ± 0.042 vs 0.075 ± 0.010,0.073 ± 0.072) were significantly upregulated.CONCLUSION:HMGA2 shRNA gene silencing induces apoptosis and suppresses proliferation of MKN-45 cells.
文摘Objective:To explore the relationship of plasma homocysteine(Hcy),soluble intercellular adhesion molecule-1(sICAM-1)and high mobility group box 1 protein(HMGB1)with carotid intima-media thickness(c-IMT)in elderly patients with type 2 diabetes mellitus.Methods:A total of 100 elderly patients who were diagnosed as type 2 diabetes mellitus in Baogang Hospital of Inner Mongolia from June 2017 to May 2020 were chosen as research objects.According to c-IMT,they were divided into the normal group(n=35),the mild to moderate group(n=41)and the severe group(n=24).The expression levels of plasma Hcy,sICAM-1 and HMGB1 were compared between groups respectively.Pearson’s correlation coefficient was used to analyze the relationship of plasma Hcy,sICAM-1,HMGB1 with c-IMT.Results:The comparison in plasma Hcy,sICAM-1,HMGB1 and c-IMT among the three groups of patients was of statistical significance(p<.05).The results of correlation analysis showed that the expression levels of plasma Hcy,sICAM-1 and HMGB1 were positively correlated with c-IMT in elderly patients with type 2 diabetes mellitus(r=.627,.598,.614;p<.05).Conclusions:The expression levels of plasma Hcy,sICAM-1 and HMGB1 are abnormally increased in elderly patients with type 2 diabetes mellitus,and related to c-IMT,which can provide a strong evidence for clinical diagnosis and treatment by detecting their levels in clinical practice.
文摘目的探讨High mobility group A2protein(HMGA2)的表达对非小细胞肺癌生长、转移的影响,与临床病理参数和细胞增殖的关系。方法应用免疫组化法检测38例非小细胞肺癌组织(23例鳞癌,15例腺癌)及癌旁的正常肺组织标本中HMGA2和Ki-67的表达。结果HMGA2和Ki-67在癌旁的正常肺组织中均不表达,而在肺癌组织中的表达阳性率分别为39.47%、44.74%,二者在肺癌组和癌旁的正常肺组织组之间差异有显著性(P<0.05)。HMGA2的表达在伴淋巴结转移的肺癌组织明显高于不伴有淋巴结转移的肺癌组织(P<0.05),而与其他临床病理参数包括肿瘤的分化程度、肿瘤大小和TNM分期以及细胞增殖指标Ki-67之间没有相关性。结论本研究显示HMGA2在非小细胞肺癌组织中异常表达,可能与肺癌的发生和进展有关。
基金supported by National Natural Science Foundation of China(6821183728)
文摘Objective:To analyze effects of high mobility group AT-hook 2(HMGA2)on malignant degree,invasion,metastasis,proliferation and cellular morphology of ovarian cancer cells.Methods:Three methods were applied to observe the effect on HMGA2 expression in ovarian cancer cells and ovarian epithelial cells.Results:After the application of siRNA-HMGA2,number of T29A2-cell clones was decreased,there was significant difference compared with the negative control Block-iT.After application of let-7c,number of T29A2+cell clones was decreased significantly,however,after the application of Anti-let-7,the number of clones restored,and there was no significant difference compared with the negative control group.After interference,the number of T29A2-cells which passed through Matrigel polycarbonate membrane were significantly lower than the negative control group.After the treatment of siRNA-HMGA2,let-7c and sh-HMGA2respectively,growth and proliferation of T29A2-,T29A2+and SKOV3 were slower,and the phenomenon was most obvious in SKOV3.Stable interference of HMGA2 induced mesenchymalepithelial changes in the morphology of SKOV3-sh-HMGA2.Conclusions:HMGA2 can promote malignant transformation of ovarian cancer cells,enhance cell invasion and metastasis,and promote cell growth and proliferation of ovarian cancer cells,which can cause ovarian cancer to progress rapidly and affect the quality of life.
文摘High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression occurrs in many kinds of cancers including colorectal cancer,suggesting that HMGA2 might play a critical role in the progression of various tumors.However,the exact role of HMGA2 in colorectal cancer has not been determined.To verify the essential role of HMGA2 in the growth and invasiveness of colorectal cancer,HMGA2 expression was down-regulated by RNA interference（RNAi） in SW480 cells.We observed that the knockdown of HMGA2 led to the significant inhibition of proliferation and invasion of SW480 cells in vitro.These results suggest that HMGA2 might play a crucial role in the progression of colorectal cancer,and be a potential therapeutic target for human colorectal cancer.
