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Objective To explore the effect of Src on cervical cancer cells through ERK signal transduction pathway.Methods Experimental study was carried out in vitro.Cervical cancer cell lines Hela (HPV-positive) and C33A (HPV-negative) were treated with Src kinase inhibitor PP2.Then,the cell cycle and apoptosis of each group were evaluated by using flow cytometry (FCM).Western blotting and Real-time PCR were used to detect the levels of the expression of ERK 1/2,c-Fos and c-Jun mRNA and protein respectively.The database was established and analyzed with SPSS statistical software (version 20.0).Results After down-regulating Src,the cell proliferation was inhibited and cell apoptosis was induced.The proportions of G0/G 1 stage of Hela and C33A cell in cell cycle increased while G2/M and S stages decreased.Meanwhile,the mRNA levels of ERK 1,ERK 2,c-Fos and c-Jun increased.And the expression levels of ERK 1/2,phosphorylated ERK 1/2 (p-ERK 1/2)and phosphorylated c-Fos (p-c-Fos) protein decreased,while c-Jun and phosphorylated c-Jun (p-c-Jun)protein expression increased.In addtion,the change level of Hela cell,p-ERK 1/2 and c-Fos protein were lower than that of C33A cell before and after the Src inhibition.Conclusions Src,involved in regulating the expression of key factors of the ERK signal transduction pathway including p-ERK 1/2 and p-c-Fos,might be capable of promoting the proliferation of cervical cancer cells and inhibiting their apoptosis.The infection with HPV might have adjustable effect on this process.
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Objective To explore the effect ofpolycyclic aromatic hydrocarbons (PAHs) and p16,FHIT gene CpG island methylation,as well as their interaction in cervical intraepithelial neoplasias.Methods Objects of this study were from a cohort of cervical lesions study in Yangqu county of Shanxi province.All the patients were diagnosed pathologically,that including 83 patients with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),86 patients with low-grade cervical intraepithelial neoplasia (C1N Ⅰ) and another 91 women under normal cervical (NC) condition.1-hydroxy pyrene in the urine was detected by high performance liquid chromatography (HPLC)while CpG island methylation status of tumor suppressor gene p16 and FHIT were measured by methylation-specifc polymerase chain reaction (MSP).Data were analyzed with Kruskal-Wallis H test,chi-square test and trend of chi-square test.Logistic regression models were used to estimate the odds ratio (OR) and corresponding 95％ confidence intervals (95％CI) between influencing factors and the cervical disease by using the SPSS statistical software (version 20.0).The interaction under study was evaluated by using the generalized multifactor dimensionality reduction (GMDR) model.Results Level of 1-hydroxy pyrene (H=50.743,P＜0.001) and the high exposure rate of 1-hydroxy pyrene (trend x2=20.146,P＜0.001) were gradually increasing along with the severity of cervical intraepithelial neoplasia.The CpG island methylation rates ofpl6,FHIT in CIN] and CIN Ⅱ/Ⅲl group were higher than that in NC group,and gradually increasing along with the severity of cervical intraepithelial neoplasia (trend x2=9.75,P=0.002;trend x 2 =10.39,P=0.001).Results from the GMDR model showed that interaction existed among the high exposure of l-hydroxy pyrene and the CpG island methylation ofpl6,FHIT in CIN Ⅰ and CIN Ⅱ]/Ⅲ group.Conclusion Under the high exposure of 1-hydroxy pyrene and the CpG island methylation of p16,FHIT appeared to have increased the risk of cervical intraepithelial neoplasia and causing synergistic effect in cervical intraepithelial neoplasia.
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Objective To explore the effect of Src on cervical cancer cells through ERK signal transduction pathway.Methods Experimental study was carried out in vitro.Cervical cancer cell lines Hela (HPV-positive) and C33A (HPV-negative) were treated with Src kinase inhibitor PP2.Then,the cell cycle and apoptosis of each group were evaluated by using flow cytometry (FCM).Western blotting and Real-time PCR were used to detect the levels of the expression of ERK 1/2,c-Fos and c-Jun mRNA and protein respectively.The database was established and analyzed with SPSS statistical software (version 20.0).Results After down-regulating Src,the cell proliferation was inhibited and cell apoptosis was induced.The proportions of G0/G 1 stage of Hela and C33A cell in cell cycle increased while G2/M and S stages decreased.Meanwhile,the mRNA levels of ERK 1,ERK 2,c-Fos and c-Jun increased.And the expression levels of ERK 1/2,phosphorylated ERK 1/2 (p-ERK 1/2)and phosphorylated c-Fos (p-c-Fos) protein decreased,while c-Jun and phosphorylated c-Jun (p-c-Jun)protein expression increased.In addtion,the change level of Hela cell,p-ERK 1/2 and c-Fos protein were lower than that of C33A cell before and after the Src inhibition.Conclusions Src,involved in regulating the expression of key factors of the ERK signal transduction pathway including p-ERK 1/2 and p-c-Fos,might be capable of promoting the proliferation of cervical cancer cells and inhibiting their apoptosis.The infection with HPV might have adjustable effect on this process.
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Objective To explore the effect ofpolycyclic aromatic hydrocarbons (PAHs) and p16,FHIT gene CpG island methylation,as well as their interaction in cervical intraepithelial neoplasias.Methods Objects of this study were from a cohort of cervical lesions study in Yangqu county of Shanxi province.All the patients were diagnosed pathologically,that including 83 patients with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),86 patients with low-grade cervical intraepithelial neoplasia (C1N Ⅰ) and another 91 women under normal cervical (NC) condition.1-hydroxy pyrene in the urine was detected by high performance liquid chromatography (HPLC)while CpG island methylation status of tumor suppressor gene p16 and FHIT were measured by methylation-specifc polymerase chain reaction (MSP).Data were analyzed with Kruskal-Wallis H test,chi-square test and trend of chi-square test.Logistic regression models were used to estimate the odds ratio (OR) and corresponding 95％ confidence intervals (95％CI) between influencing factors and the cervical disease by using the SPSS statistical software (version 20.0).The interaction under study was evaluated by using the generalized multifactor dimensionality reduction (GMDR) model.Results Level of 1-hydroxy pyrene (H=50.743,P＜0.001) and the high exposure rate of 1-hydroxy pyrene (trend x2=20.146,P＜0.001) were gradually increasing along with the severity of cervical intraepithelial neoplasia.The CpG island methylation rates ofpl6,FHIT in CIN] and CIN Ⅱ/Ⅲl group were higher than that in NC group,and gradually increasing along with the severity of cervical intraepithelial neoplasia (trend x2=9.75,P=0.002;trend x 2 =10.39,P=0.001).Results from the GMDR model showed that interaction existed among the high exposure of l-hydroxy pyrene and the CpG island methylation ofpl6,FHIT in CIN Ⅰ and CIN Ⅱ]/Ⅲ group.Conclusion Under the high exposure of 1-hydroxy pyrene and the CpG island methylation of p16,FHIT appeared to have increased the risk of cervical intraepithelial neoplasia and causing synergistic effect in cervical intraepithelial neoplasia.
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Objective To explore the effects of both folic acid,p16 protein expression and their interaction on progression of cervical cancerization.Methods Participants were pathologically diagnosed new cases,including 80 women with normal cervical (NC),55 patients with low-grade cervical intraepithelial neoplasia (CIN Ⅰ),55 patients with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ) and 64 patients with cervical squamous cell carcinoma (SCC).Serum folate levels were detected by microbiological assay method while p16 protein expression levels were measured by Western-blot.In vitro,cervical cancer cell lines C33A (HPV negative) and Caski (HPV16 positive) were treated with different concentrations of folate.Proliferation and apoptosis of cells and the levels of p16 protein expression were measured in groups with different folic acid concentrations.Results Results showed that the levels of serum folate were (5.96± 3.93) ng/ml,(5.08±3.43) ng/ml,(3.92 ± 2.59) ng/ml and (3.18 ± 2.71) ng/ml,and the levels ofpl6 protein were 0.80 ± 0.32,1.33 ± 0.52,1.91 ± 0.77,and 2.09 ± 0.72 in the group of NC,CIN Ⅰ,CIN Ⅱ/Ⅲ and SCC,respectively.However,the levels of serum folate decreased (trend X2 =32.71,P＜ 0.001) and p 16 protein expression increased (trend x2=56.06,P＜0.001) gradually along with the severity of cervix lesions.An additive interaction was seen between serum folate deficiency and high expression of p l 6 protein in the CIN Ⅰ,CIN n/Ⅲ and SCC group.Results in vitro showed that,with the increase of folate concentration,the inhibition rate of cell proliferation (C33A:r=0.928,P=0.003;Caski:r=0.962,P=0.001) and the rate on cell apoptosis (C33A:r=0.984,P＜0.001;Caski:r=0.986,P＜0.001) all increased but the levels of p16 protein expression (C33A:r=-0.817,P=0.025;Caski:r=-0.871,P=0.011) reduced.The proliferation inhibition rate (C33A:r=-0.935,P=0.002;Caski:r=-0.963,P=0.001) and apoptosis rate of cells (C33A:r=-0.844,P=0.017;Caski:r=-0.898,P=0.006) were negatively correlated with the levels of p16 protein expression.Conclusions Our findings indicated that both serum folate deficiency and high expression of p16 protein could increase the risk of cervical cancer and cervix precancerous lesion,and there was an additive interaction between them.Our findings suggested that folic acid supplementation could reverse the abnormal expression of p16 protein,and effectively promote apoptosis and inhibit proliferation in cervical carcinoma cells.
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Objective To explore the effects of both folic acid,p16 protein expression and their interaction on progression of cervical cancerization.Methods Participants were pathologically diagnosed new cases,including 80 women with normal cervical (NC),55 patients with low-grade cervical intraepithelial neoplasia (CIN Ⅰ),55 patients with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ) and 64 patients with cervical squamous cell carcinoma (SCC).Serum folate levels were detected by microbiological assay method while p16 protein expression levels were measured by Western-blot.In vitro,cervical cancer cell lines C33A (HPV negative) and Caski (HPV16 positive) were treated with different concentrations of folate.Proliferation and apoptosis of cells and the levels of p16 protein expression were measured in groups with different folic acid concentrations.Results Results showed that the levels of serum folate were (5.96± 3.93) ng/ml,(5.08±3.43) ng/ml,(3.92 ± 2.59) ng/ml and (3.18 ± 2.71) ng/ml,and the levels ofpl6 protein were 0.80 ± 0.32,1.33 ± 0.52,1.91 ± 0.77,and 2.09 ± 0.72 in the group of NC,CIN Ⅰ,CIN Ⅱ/Ⅲ and SCC,respectively.However,the levels of serum folate decreased (trend X2 =32.71,P＜ 0.001) and p 16 protein expression increased (trend x2=56.06,P＜0.001) gradually along with the severity of cervix lesions.An additive interaction was seen between serum folate deficiency and high expression of p l 6 protein in the CIN Ⅰ,CIN n/Ⅲ and SCC group.Results in vitro showed that,with the increase of folate concentration,the inhibition rate of cell proliferation (C33A:r=0.928,P=0.003;Caski:r=0.962,P=0.001) and the rate on cell apoptosis (C33A:r=0.984,P＜0.001;Caski:r=0.986,P＜0.001) all increased but the levels of p16 protein expression (C33A:r=-0.817,P=0.025;Caski:r=-0.871,P=0.011) reduced.The proliferation inhibition rate (C33A:r=-0.935,P=0.002;Caski:r=-0.963,P=0.001) and apoptosis rate of cells (C33A:r=-0.844,P=0.017;Caski:r=-0.898,P=0.006) were negatively correlated with the levels of p16 protein expression.Conclusions Our findings indicated that both serum folate deficiency and high expression of p16 protein could increase the risk of cervical cancer and cervix precancerous lesion,and there was an additive interaction between them.Our findings suggested that folic acid supplementation could reverse the abnormal expression of p16 protein,and effectively promote apoptosis and inhibit proliferation in cervical carcinoma cells.
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Objective To study a scoring method for assessing function status of cardiovascular system quantitatively. Methods First, exercise heart rate variability (EHRV) was obtained from the primary dynamic electrocardiogram (ECG) recorded during exercise testing. Then appropriate entropy, relative complexity, and other three parameters extracted from poincare dispersed-dot plot were extracted from the EHRV. Discriminant analysis was used to classify two extreme groups.Results Based on the values of two groups from the discriminant formula, a scoring formula was proposed and four ranks were divided according to different score domains. A novel scoring method was established. To validate the proposed scoring method, 60 middle-aged hypertensives, 50 elder subjects and 110 young healthy subjects were examined and scored. Conclusion Scores of 220 subjects are consistent with their real health status. The proposed scoring method reflects the status of the subject's cardiovascular system effectively.