BACKGROUND Cancerous inhibitor of protein phosphatase 2A(CIP2A)is a newly discovered oncogene.It is an active cell proliferation regulatory factor that inhibits tumor apoptosis in gastric cancer(GC)cells.CIP2A is func...BACKGROUND Cancerous inhibitor of protein phosphatase 2A(CIP2A)is a newly discovered oncogene.It is an active cell proliferation regulatory factor that inhibits tumor apoptosis in gastric cancer(GC)cells.CIP2A is functionally related to chemoresistance in various types of tumors according to recent studies.The underlying mechanism,however,is unknown.Further,the primary treatment regimen for GC is oxaliplatin-based chemotherapy.Nonetheless,it often fails due to chemoresistance of GC cells to oxaliplatin.AIM The goal of this study was to examine CIP2A expression and its association with oxaliplatin resistance in human GC cells.METHODS Immunohistochemistry was used to examine CIP2A expression in GC tissues and adjacent normal tissues.CIP2A expression in GC cell lines was reduced using small interfering RNA.After confirming the silencing efficiency,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium and flow cytometry assays were used to evaluate cell proliferation and apoptosis caused by oxaliplatin treatment.Further,the key genes and protein changes were verified using realtime quantitative reverse transcription PCR and Western blotting,respectively,before and after intervention.For bioinformatics analysis,we used the R software and Bioconductor project.For statistical analysis,we used GraphPad Prism 6.0 and the Statistical Package for the Social Sciences software version 20.0(IBM,Armonk,United States).RESULTS A high level of CIP2A expression was associated with tumor size,T stage,lymph node metastasis,Tumor Node Metastasis stage,and a poor prognosis.Further,CIP2A expression was higher in GC cells than in normal human gastric epithelial cells.Using small interfering RNA against CIP2A,we discovered that CIP2A knockdown inhibited cell proliferation and significantly increased GC cell sensitivity to oxaliplatin.Moreover,CIP2A knockdown enhanced oxaliplatin-induced apoptosis in GC cells.Hence,high CIP2A levels in GC may be a factor in chemoresistance to oxaliplatin.In human GC cells,CIP2A regulated protein kinase B phosphorylation,and chemical inhibition of the protein kinase B signaling pathway was significantly associated with increased sensitivity to oxaliplatin.Therefore,the protein kinase B signaling pathway was correlated with CIP2Aenhanced chemoresistance of human GC cells to oxaliplatin.CONCLUSION CIP2A expression could be a novel therapeutic strategy for chemoresistance in GC.展开更多
Protein phosphatase 2 A（PP2 A） was reported to play an important role in cancer development;however,the relationship between PP2 A and cervical cancer development has yet to be fully understood.The present study aim...Protein phosphatase 2 A（PP2 A） was reported to play an important role in cancer development;however,the relationship between PP2 A and cervical cancer development has yet to be fully understood.The present study aimed to explore the role of PP2 A in the development of cervical cancer.Serum levels of PP2 A were detected by ELISA in 23 patients with cervical cancer and 30 patients with benign cervical lesions.Furthermore,the PP2 A activities and the m RNA and protein levels of PP2 A were measured in cervical cancer（n=8） and chronic cervicitis（n=10） tissues.The results showed that the serum levels of PP2 A were significantly reduced in patients with cervical cancer.Further studies showed that not only the activities of PP2 A but also the m RNA and protein levels of PP2 A were significantly decreased in cervical cancer tissues.Wound healing and Transwell assays demonstrated that pharmacological and genetic upregulation of PP2 A could inhibit the migration of He La cells,but the downregulation of PP2 A promoted cellular migration.The activation of PP2 A also inhibited the remodeling of actin and the activity of mitogen-activated protein kinases（MAPKs） including p-JNK,p-p38 and p-ERK.Meanwhile,the activation of PP2 A was found to downregulate MMP-9 levels,which further inhibited the migration and invasion of He La cells.In conclusion,our data suggest that the activity and expression of PP2 A are significantly reduced in cervical cancer tissues,and the activation of PP2 A may inhibit the migration of cervical cancer cells by inhibiting the phosphorylation of p-JNK,p-p38 and the p-ERK/MAPK signaling pathway as well as by downregulating MMP-9,implying that PP2 A plays an important role in cervical cancer development.展开更多
AIM To investigate the role of calmodulin-dependent protein kinase Ⅱ(Ca MKⅡ) in colon cancer growth,migration and invasion.METHODS Ca MKⅡ expression in colon cancer and paracancerous tissues was evaluated via immun...AIM To investigate the role of calmodulin-dependent protein kinase Ⅱ(Ca MKⅡ) in colon cancer growth,migration and invasion.METHODS Ca MKⅡ expression in colon cancer and paracancerous tissues was evaluated via immunochemistry. Transcriptional and posttranscriptional levels of Ca MKⅡin tissue samples and MMP2,MMP9 and TIMP-1 expression in the human colon cancer cell line HCT116 were assessed by q RTPCR and western blot. Cell proliferation was detected with the MTT assay. Cancer cell migration and invasion were investigated with the Transwell culture system and woundhealing assay.RESULTS We first demonstrated that CaMK Ⅱ was ove rexpressed in human colon cancers and was associated with cancer differentiation. In the human colon cancer cell line HCT116,the Ca MKII-specific inhibitor KN93,but not its inactive analogue KN92,decreased cancer cell proliferation. Furthermore,KN93 also significantly prohibited HCT116 cell migration and invasion. The specific inhibition of ERK1/2 or p38 decreased the proliferation and migration of colon cancer cells.CONCLUSION Our findings highlight Ca MKⅡ as a potential critical mediator in human colon tumor development and metastasis.展开更多
This study investigated the expression and prognostic value of SHP-2 in cervical cancer caused by human papillomavirus (HPV) infection. Forty-five specimens from patients with cervical cancer (stageⅠ-Ⅲ), 32 specimen...This study investigated the expression and prognostic value of SHP-2 in cervical cancer caused by human papillomavirus (HPV) infection. Forty-five specimens from patients with cervical cancer (stageⅠ-Ⅲ), 32 specimens from patients with cervical intraepithelial neoplasia (CIN) (Ⅰ, Ⅱ) and 20 normal cervical samples from patients with hysteromyoma were collected in Department of Pathology for comparison. The expression levels of SHP-2 and IFN-β proteins were detected by using immunohistochemistry. The mRNA expression level of SHP-2 was detected by using quantitative real-time polymerase chain reaction (PCR). HPVs were detected by HPV GenoArray Test. The Spearman correlation was used to compare the expression level of SHP-2 in HPV infected cervical cancer vs non-HPV infected normal cervix. The level of SHP-2 protein expression in the cancer tissues (88.8%) was significantly higher than in CIN tissues (62.5%) and normal cervixes (45%) (P<0.05 and P<0.05, respectively). The SHP-2 mRNA levels in the cancer tissues were upregulated as compared with those in the normal cervixes (P<0.05). Twenty-one (46.7%) cervical cancers, 25 (78.1%) CINs and 17 (85%) normal cervixes showed IFN-β positive staining in cytoplasm. There was statistically significant difference in the expression rate of IFN-β between cervical cancer and normal cervix (χ2=8.378, P<0.05) as well as between cervical cancer and CIN (χ2=7.695, P<0.05). HPV16/18 infections could be found in normal cervixs (15%), CINs (68.7%) and cervical cancers (84.4%). There was a correlation between HPV infection and SHP-2 expression in cervical cancer (rs=0.653, P<0.05). SHP-2 may be a useful prognostic and diagnostic indicator for HPV infected cervical cancer. In cervical cancers, SHP-2 mRNA and protein overexpression was associated with IFN-β lower-expression.展开更多
AIM:To explore the alteration of tyrosine phosphatase SHP-2 protein expression in gastric cancer and to assess its prognostic values.METHODS:Three hundred and five consecutive cases of gastric cancer were enrolled int...AIM:To explore the alteration of tyrosine phosphatase SHP-2 protein expression in gastric cancer and to assess its prognostic values.METHODS:Three hundred and five consecutive cases of gastric cancer were enrolled into this study.SHP-2 expression was carried out in 305 gastric cancer specimens,of which 83 were paired adjacent normal gastric mucus samples,using a tissue microarray immunohistochemical method.Correlations were analyzed between expression levels of SHP-2 protein and tumor parameters or clinical outcomes.Serum anti-Helicobacter pylori(H.pylori) immunoglobulin G was detected with enzyme-linked immunosorbent assay.Cox proportional hazards model was used to evaluate prognostic values by compassion of the expression levels of SHP-2 and disease-specific survivals in patients.RESULTS:SHP-2 staining was found diffuse mainly in the cytoplasm and the weak staining was also observed in the nucleus in gastric mucosa cells.Thirty-two point five percent of normal epithelial specimen and 62.6% of gastric cancer specimen were identified to stain with SHP-2 antibody positively(P < 0.001).Though SHP-2 staining intensities were stronger in the H.pylori(+) group than in the H.pylori(-) group,no statistically significant difference was found in the expression levels of SHP-2 between H.pylori(+) and H.pylori(-) gastric cancer(P = 0.40).The SHP-2 expression in gastric cancer was not significantly associated with cancer stages,lymph node metastases,and distant metastasis of the tumors(P = 0.34,P = 0.17,P = 0.52).Multivariate analysis demonstrated no correlation between SHP-2 expression and disease-free survival(P = 0.86).CONCLUSION:Increased expression of SHP-2 protein in gastric cancer specimen suggesting the aberrant upregulation of SHP-2 protein might play an important role in the gastric carcinogenesis.展开更多
Objective: To study the correlation of SFRP2 and TPX2 expression with cancer cell apoptosis and EMT process in cervical cancer. Methods: Patients with cervical cancer who received surgical resection in No.174 Hospital...Objective: To study the correlation of SFRP2 and TPX2 expression with cancer cell apoptosis and EMT process in cervical cancer. Methods: Patients with cervical cancer who received surgical resection in No.174 Hospital of PLA between June 2015 and December 2016 were selected as the research subjects, moderate amount of cervical cancer lesion and adjacent lesion were collected after surgical resection to extract RNA, and then fluorescent quantitative PCR kit was used to determine the expression of SFRP2, TPX2, apoptosis genes and EMT genes in lesion tissue. Results: SFRP2, SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression in cervical cancer lesion were greatly lower than those in adjacent lesions whereas TPX2, SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression were greatly higher than those in adjacent lesions;SFRP2 mRNA expression in cervical cancer lesion was positively correlated with SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression, and negatively correlated with SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression whereas TPX2 mRNA expression was negatively correlated with SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression, and positively correlated with SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression. Conclusion: The lowly expressed SFRP2 and highly expressed TPX2 in cervical cancer lesions can inhibit the apoptosis of cancer cells and promote the EMT process of cancer cells.展开更多
Our preliminary proteomics analysis suggested that expression of microtubule-associated protein tau is elevated in the spinal cord after injury. Therefore, the first aim of the present study was to examine tau express...Our preliminary proteomics analysis suggested that expression of microtubule-associated protein tau is elevated in the spinal cord after injury. Therefore, the first aim of the present study was to examine tau expression in the injured spinal cord. The second aim was to determine whether tau can regulate neural stem cell migration, a critical factor in the successful treatment of spinal cord injury. We established rat models of spinal cord injury and injected them with mouse hippocampal neural stem cells through the tail vein. We used immunohistochemistry to show that the expression of tau protein and the number of migrated neural stem cells were markedly increased in the injured spinal cord. Furthermore, using a Transwell assay, we showed that neural stem cell migration was not affected by an elevated tau concentration in the outer chamber, but it was decreased by changes in intracellular tau phosphorylation state. These results demonstrate that neural stem cells have targeted migration capability at the site of injury, and that although tau is not a chemokine for targeted migration of neural stem cells, intracellular tau phosphorylation/dephosphorylation can inhibit cell migration.展开更多
基金Supported by This work was supported by the Natural Science Foundation of Gansu Province,China,No.17JR5RA272 and No.22JR5RA923the Research Fund Project of The First Hospital of Lanzhou University,No.ldyyyn2021-120,No.ldyyyn2020-98 and No.ldyyyn2021-30.
文摘BACKGROUND Cancerous inhibitor of protein phosphatase 2A(CIP2A)is a newly discovered oncogene.It is an active cell proliferation regulatory factor that inhibits tumor apoptosis in gastric cancer(GC)cells.CIP2A is functionally related to chemoresistance in various types of tumors according to recent studies.The underlying mechanism,however,is unknown.Further,the primary treatment regimen for GC is oxaliplatin-based chemotherapy.Nonetheless,it often fails due to chemoresistance of GC cells to oxaliplatin.AIM The goal of this study was to examine CIP2A expression and its association with oxaliplatin resistance in human GC cells.METHODS Immunohistochemistry was used to examine CIP2A expression in GC tissues and adjacent normal tissues.CIP2A expression in GC cell lines was reduced using small interfering RNA.After confirming the silencing efficiency,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium and flow cytometry assays were used to evaluate cell proliferation and apoptosis caused by oxaliplatin treatment.Further,the key genes and protein changes were verified using realtime quantitative reverse transcription PCR and Western blotting,respectively,before and after intervention.For bioinformatics analysis,we used the R software and Bioconductor project.For statistical analysis,we used GraphPad Prism 6.0 and the Statistical Package for the Social Sciences software version 20.0(IBM,Armonk,United States).RESULTS A high level of CIP2A expression was associated with tumor size,T stage,lymph node metastasis,Tumor Node Metastasis stage,and a poor prognosis.Further,CIP2A expression was higher in GC cells than in normal human gastric epithelial cells.Using small interfering RNA against CIP2A,we discovered that CIP2A knockdown inhibited cell proliferation and significantly increased GC cell sensitivity to oxaliplatin.Moreover,CIP2A knockdown enhanced oxaliplatin-induced apoptosis in GC cells.Hence,high CIP2A levels in GC may be a factor in chemoresistance to oxaliplatin.In human GC cells,CIP2A regulated protein kinase B phosphorylation,and chemical inhibition of the protein kinase B signaling pathway was significantly associated with increased sensitivity to oxaliplatin.Therefore,the protein kinase B signaling pathway was correlated with CIP2Aenhanced chemoresistance of human GC cells to oxaliplatin.CONCLUSION CIP2A expression could be a novel therapeutic strategy for chemoresistance in GC.
基金supported by the National Natural Science Foundation of China(No.81100959)Natural Science Foundation of Hubei Province(Nos.2015CFB185,2015CFB723)National Clinical Key Specialty Construction Projects of Ministry of Health of China(No.2010-305)
文摘Protein phosphatase 2 A（PP2 A） was reported to play an important role in cancer development;however,the relationship between PP2 A and cervical cancer development has yet to be fully understood.The present study aimed to explore the role of PP2 A in the development of cervical cancer.Serum levels of PP2 A were detected by ELISA in 23 patients with cervical cancer and 30 patients with benign cervical lesions.Furthermore,the PP2 A activities and the m RNA and protein levels of PP2 A were measured in cervical cancer（n=8） and chronic cervicitis（n=10） tissues.The results showed that the serum levels of PP2 A were significantly reduced in patients with cervical cancer.Further studies showed that not only the activities of PP2 A but also the m RNA and protein levels of PP2 A were significantly decreased in cervical cancer tissues.Wound healing and Transwell assays demonstrated that pharmacological and genetic upregulation of PP2 A could inhibit the migration of He La cells,but the downregulation of PP2 A promoted cellular migration.The activation of PP2 A also inhibited the remodeling of actin and the activity of mitogen-activated protein kinases（MAPKs） including p-JNK,p-p38 and p-ERK.Meanwhile,the activation of PP2 A was found to downregulate MMP-9 levels,which further inhibited the migration and invasion of He La cells.In conclusion,our data suggest that the activity and expression of PP2 A are significantly reduced in cervical cancer tissues,and the activation of PP2 A may inhibit the migration of cervical cancer cells by inhibiting the phosphorylation of p-JNK,p-p38 and the p-ERK/MAPK signaling pathway as well as by downregulating MMP-9,implying that PP2 A plays an important role in cervical cancer development.
基金Supported by the National Natural Science Foundation of China,No.81302131
文摘AIM To investigate the role of calmodulin-dependent protein kinase Ⅱ(Ca MKⅡ) in colon cancer growth,migration and invasion.METHODS Ca MKⅡ expression in colon cancer and paracancerous tissues was evaluated via immunochemistry. Transcriptional and posttranscriptional levels of Ca MKⅡin tissue samples and MMP2,MMP9 and TIMP-1 expression in the human colon cancer cell line HCT116 were assessed by q RTPCR and western blot. Cell proliferation was detected with the MTT assay. Cancer cell migration and invasion were investigated with the Transwell culture system and woundhealing assay.RESULTS We first demonstrated that CaMK Ⅱ was ove rexpressed in human colon cancers and was associated with cancer differentiation. In the human colon cancer cell line HCT116,the Ca MKII-specific inhibitor KN93,but not its inactive analogue KN92,decreased cancer cell proliferation. Furthermore,KN93 also significantly prohibited HCT116 cell migration and invasion. The specific inhibition of ERK1/2 or p38 decreased the proliferation and migration of colon cancer cells.CONCLUSION Our findings highlight Ca MKⅡ as a potential critical mediator in human colon tumor development and metastasis.
基金supported by a grant from the National Natu-ral Science Foundation of China(No.30973191)
文摘This study investigated the expression and prognostic value of SHP-2 in cervical cancer caused by human papillomavirus (HPV) infection. Forty-five specimens from patients with cervical cancer (stageⅠ-Ⅲ), 32 specimens from patients with cervical intraepithelial neoplasia (CIN) (Ⅰ, Ⅱ) and 20 normal cervical samples from patients with hysteromyoma were collected in Department of Pathology for comparison. The expression levels of SHP-2 and IFN-β proteins were detected by using immunohistochemistry. The mRNA expression level of SHP-2 was detected by using quantitative real-time polymerase chain reaction (PCR). HPVs were detected by HPV GenoArray Test. The Spearman correlation was used to compare the expression level of SHP-2 in HPV infected cervical cancer vs non-HPV infected normal cervix. The level of SHP-2 protein expression in the cancer tissues (88.8%) was significantly higher than in CIN tissues (62.5%) and normal cervixes (45%) (P<0.05 and P<0.05, respectively). The SHP-2 mRNA levels in the cancer tissues were upregulated as compared with those in the normal cervixes (P<0.05). Twenty-one (46.7%) cervical cancers, 25 (78.1%) CINs and 17 (85%) normal cervixes showed IFN-β positive staining in cytoplasm. There was statistically significant difference in the expression rate of IFN-β between cervical cancer and normal cervix (χ2=8.378, P<0.05) as well as between cervical cancer and CIN (χ2=7.695, P<0.05). HPV16/18 infections could be found in normal cervixs (15%), CINs (68.7%) and cervical cancers (84.4%). There was a correlation between HPV infection and SHP-2 expression in cervical cancer (rs=0.653, P<0.05). SHP-2 may be a useful prognostic and diagnostic indicator for HPV infected cervical cancer. In cervical cancers, SHP-2 mRNA and protein overexpression was associated with IFN-β lower-expression.
基金Supported by National Natural Science Foundation of China,No. 81072369Natural Science Foundation of Jilin Province,China,No. 200905131
文摘AIM:To explore the alteration of tyrosine phosphatase SHP-2 protein expression in gastric cancer and to assess its prognostic values.METHODS:Three hundred and five consecutive cases of gastric cancer were enrolled into this study.SHP-2 expression was carried out in 305 gastric cancer specimens,of which 83 were paired adjacent normal gastric mucus samples,using a tissue microarray immunohistochemical method.Correlations were analyzed between expression levels of SHP-2 protein and tumor parameters or clinical outcomes.Serum anti-Helicobacter pylori(H.pylori) immunoglobulin G was detected with enzyme-linked immunosorbent assay.Cox proportional hazards model was used to evaluate prognostic values by compassion of the expression levels of SHP-2 and disease-specific survivals in patients.RESULTS:SHP-2 staining was found diffuse mainly in the cytoplasm and the weak staining was also observed in the nucleus in gastric mucosa cells.Thirty-two point five percent of normal epithelial specimen and 62.6% of gastric cancer specimen were identified to stain with SHP-2 antibody positively(P < 0.001).Though SHP-2 staining intensities were stronger in the H.pylori(+) group than in the H.pylori(-) group,no statistically significant difference was found in the expression levels of SHP-2 between H.pylori(+) and H.pylori(-) gastric cancer(P = 0.40).The SHP-2 expression in gastric cancer was not significantly associated with cancer stages,lymph node metastases,and distant metastasis of the tumors(P = 0.34,P = 0.17,P = 0.52).Multivariate analysis demonstrated no correlation between SHP-2 expression and disease-free survival(P = 0.86).CONCLUSION:Increased expression of SHP-2 protein in gastric cancer specimen suggesting the aberrant upregulation of SHP-2 protein might play an important role in the gastric carcinogenesis.
文摘Objective: To study the correlation of SFRP2 and TPX2 expression with cancer cell apoptosis and EMT process in cervical cancer. Methods: Patients with cervical cancer who received surgical resection in No.174 Hospital of PLA between June 2015 and December 2016 were selected as the research subjects, moderate amount of cervical cancer lesion and adjacent lesion were collected after surgical resection to extract RNA, and then fluorescent quantitative PCR kit was used to determine the expression of SFRP2, TPX2, apoptosis genes and EMT genes in lesion tissue. Results: SFRP2, SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression in cervical cancer lesion were greatly lower than those in adjacent lesions whereas TPX2, SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression were greatly higher than those in adjacent lesions;SFRP2 mRNA expression in cervical cancer lesion was positively correlated with SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression, and negatively correlated with SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression whereas TPX2 mRNA expression was negatively correlated with SARI, AIF, FHIT, NDRG4, MCPH1 and E-cadherin mRNA expression, and positively correlated with SP2, CyclinD1, Piwil2, HERC4, EFEMP1, EZH2, TGF-β1, N-cadherin and Vimentin1 mRNA expression. Conclusion: The lowly expressed SFRP2 and highly expressed TPX2 in cervical cancer lesions can inhibit the apoptosis of cancer cells and promote the EMT process of cancer cells.
基金supported by the National Natural Science Foundation of China,No.81250016,31572217
文摘Our preliminary proteomics analysis suggested that expression of microtubule-associated protein tau is elevated in the spinal cord after injury. Therefore, the first aim of the present study was to examine tau expression in the injured spinal cord. The second aim was to determine whether tau can regulate neural stem cell migration, a critical factor in the successful treatment of spinal cord injury. We established rat models of spinal cord injury and injected them with mouse hippocampal neural stem cells through the tail vein. We used immunohistochemistry to show that the expression of tau protein and the number of migrated neural stem cells were markedly increased in the injured spinal cord. Furthermore, using a Transwell assay, we showed that neural stem cell migration was not affected by an elevated tau concentration in the outer chamber, but it was decreased by changes in intracellular tau phosphorylation state. These results demonstrate that neural stem cells have targeted migration capability at the site of injury, and that although tau is not a chemokine for targeted migration of neural stem cells, intracellular tau phosphorylation/dephosphorylation can inhibit cell migration.
