Downregulation of cytokeratin 18 increases NMIIA expression and promotes breast cancer metastasis / 中国药理学通报

Aim To investigate the role of aberrant cytokeratin 18(CK18) expression in breast cancer metastasis, anrl to elucidate the mechanism by identif¬ying its target.Methods The expression of CK.18 in human breast cancer tissues and cells was determined using immunohistochemical staining and Western blot, respectively.CK18 expression in human breast cancer MCF-7 cells was effectively down-regulated by shRNA, and its effect on breast cancer metastasis was further determined by scratch wound healing assay.The co-lo- cation of CK18 and non-muscle II A ( NMIIA) in MCF-7 cells was examined using double immunofluo¬rescence staining.The effect of CK18 down-regulation on the levels of NMIIA and c-Abl-ERK signaling was quantified by Western blot.Results Lower CK18 lev¬els was found in metastatic than that in primary breast cancer tissues and in highly invasive MDA-MB-231 than that in MCF-7 cells.CK.18 down-regulation pro¬moted the wound repair ability of MCF-7 cells 72h after scratch.CK18 and NMIIA were shown to co-locate in cytoplasm of MCF-7 cells.Moreover, down-regulation of CK18 increased NMIIA expression and activated the c-Abl-ERK signaling pathway in MCF-7 cells.Con¬clusions Down-regulation of CK18 could promote me¬tastasis of breast cancer, which is related to increased NMIIA expression and the activation of c-Abl-ERK sig¬naling pathway.

Expression of M30 and M65 in celiac disease. Analytical cross-sectional study

ABSTRACT BACKGROUND: The role of villous atrophy in apoptosis, a distinctive feature of celiac disease, is a matter of controversy. The aim of this study was to determine the apoptosis rate through immunohistochemical staining for M30 and M65 in celiac disease cases. DESIGN AND SETTING: Analytical cross-sectional study in a tertiary-level center. METHODS: Duodenal biopsies from 28 treatment-naive patients with celiac disease, 16 patients with potential celiac disease, 10 patients with a gluten-free diet and 8 controls were subjected to immunohistochemical staining for the end-apoptotic marker M30 and the total cell death marker M65. H-scores were compared. Several laboratory parameters were recorded concomitantly, and at the one-year follow-up for celiac disease and potential celiac disease patients. RESULTS: There was a significant difference in H-score for M30 expression between the celiac disease, potential celiac disease and gluten-free diet groups (P = 0.009). There was no significant difference in H-score for M65 expression. There was a positive correlation between the H-score for M30 expression and the anti-tissue transglutaminase immunoglobulin A (anti-tTgIgA) and anti-tissue transglutaminase immunoglobulin G (anti-tTgIgG) levels (R = 0.285, P = 0.036; and R = 0.307, P = 0.024, respectively); and between the H-score for M65 expression and the anti-tTgIgA and anti-tTgIgG levels (R = 0.265, P = 0.053; and R=0.314, P = 0.021, respectively). There was no difference between celiac disease and potential celiac disease patients regarding the laboratory parameters selected. CONCLUSION: The rates of apoptosis and nutritional deficiencies in patients with potential celiac disease were similar to those in patients with celiac disease.

Advances in epidemiology and serum markers for the noninvasive diagnosis of nonalcoholic fatty liver disease / 中华肝脏病杂志

The global prevalence rate of nonalcoholic fatty liver disease (NAFLD) has increased year by year, and it has become the number one cause for chronic liver disease in China. In addition, the trend of NAFLD has become more pronounced and evident in female gender and younger age group. The long-term persistence of fatty liver disease may cause serious consequences. There are no accepted diagnostic criteria for diagnosing noninvasive diagnosis of NAFLD. Alpha-ketoglutarate is a newly discovered serological marker of high diagnostic value and considered the most valuable potential biomarker along with cytokeratine-18 (CK-18).

Expressions and roles of three inflammatory cytokines in chronic hepatitis B patients with nonalcoholic fatty liver disease / 中华传染病杂志

Objective To investigate the roles of cytokeratin 18 (CK18) M30 and M65,thymosin beta 4 (Tβ4) and tumor necrosis factor (TNF)-α in hepatic steatosis and development of inflammatory and fibrosis in chronic hepatitis B (CHB) patients with nonalcoholic fatty liver disease (NAFLD).Methods A total of 46 CHB patients with NAFLD and 42 CHB patients were collected.Serum CK-18 M30,M65,Tβ4 and TNF-α levels were measured by enzyme linked immunosorbent assay (ELISA) in two groups.The associations between inflammatory factors levels and biochemical or pathological indicators were analyzed.The statistical analysis was conducted by t test and chi square test of two independent samples.The correlation analysis was performed by Pearson and Logistic regression analysis.Results The mean serum CK 18 M30 level in CHB with NAFLD group was (614.48±471.43) U/L,which was significantly higher than that in CHB group (374.50±231.04) U/L (t=2.988,P＜0.01).The mean levels of CK18 M65,Tβ4 and TNF-α in CHB with NAFLD group were (369.41±262.21) U/L,(0.80±0.32) mg/L and (54.87±20.36) ng/L,respectively,and those in CHB group were (296.50±231.44) U/L,(0.68±0.30) mg/L and (51.88± 20.60) ng/L,respectively.There were no difference between CHB with NAFLD group and CHB group (t=1.378,1.810 and 0.685,respectively,all P＞0.05).In CHB with NAFLD patients,the CK-18 M30 level was positively correlated with alanine aminotransferase,triglyceride,fasting blood glucose,histology inflammation score,fibrosis score and steatosis (r=0.507,0.456,0.384,0.551,0.458 and 0.457,respectively,all P＜0.01).Tβ4 level was negatively correlated with inflammation and fibrosis score (r=0.371 and-0.308,respectively,P＜0.05).TNF-α level was positively correlated with inflammation score and steatosis (r=0.570 and 0.441,respectively,P＜0.01).CK-18 M30,Tβ4 and TNF-α were independent predictors of CHB combined with NAFLD,progressive inflammatory fibrosis and severe steatosis.Conclusions Serum CK-18 M30,Tβ4 and TNF-α levels are associated with hepatic steatosis,development of inflammation and fibrosis in CHB with NAFLD patients.

Clinical value of Serum CK-18 M30 levels in patients with nonalcoholic steatohepatitis / 实用医学杂志

Objective To detect the level of serum fragmented cytokeratin 18 (CK-18 M30) in patients with nonalcoholic steatohepatitis (NASH), to explore the relationship between the expression of CK-18 M30 and NASH. Methods 33 healthy people as control group, 24 nonalcoholic simple fatty liver (NAFL) patients, and 21 NASH patients were included in this study. CK-18 M30, ALT, AST and GGT were detected in all patients’ vein blood. NAFLD activity points (NAS) was examined in biopsy specimens of NAFL patients and NASH patients. Pearson correlation was applied to analyze the correlations between serum CK-18 M30, ALT, AST, GGT and the NAS of liver tissue in NASH group. Results Serum CK-18 M30 level of healthy control, NAFL and NASH group were (96.557 2 ± 41.226 8)U/L, (104.321 7 ± 45.167 3)U/L, (263.125 5 ± 61.578 1)U/L respectively. Serum CK-18 M30 level in NASH patients positively correlated with both NAS of liver tissue and serum ALT, which correlation coefficient r values were 0.601 5 and 0.420 6. Conclusion The concentration of serum CK-18 M30 could be used as a marker in the diagnosis of NASH.

The application value of combined detection of TuM2-PK,CK18-3A9 and CYFRA21-1 in diagnosis of NSCLC / 国际检验医学杂志

Objective To investigate the value of tumor M2 pyruvate kinase (TuM2-PK ) ,cytokeratin-18 (CK18 )-3A9 and cytokeratin 19 fragment(CYFRA21-1) in non small cell lung cancer(NSCLC) ,and to evaluate the application value of combined de-tection of these three kinds tumor markers in diagnosis of NSCLC .Methods The serum levels of TuM2-PK and CK18-3A9 were measured in 67 patients with NSCLC(NSCLC group) ,72 patients with benign lung diseases(benign group) and 75 healthy control (control group) by enzyme linked immunosorbent assay(ELISA) .The level of CYFRA21-1 was detected by electrochemilumines-cence method .Results The serum levels of TuM2-PK ,CK18-3A9 and CYFRA21-1 in NSCLC group were higher than those in be-nign group and control group(P<0 .05) .The sensitivity of TuM2-PK or CK18-3A9 was better than CYFRA21-1(P<0 .05) ,the specificity of CYFRA21-1 was better than TuM2-PK or CK18-3A9(P<0 .01) .The sensitivity and accuracy of combined detection of TuM2-PK ,CK18-3A9 and CYFRA21-1 were both better than CYFRA21-1(P<0 .05) .The sensitivity ,specificity and accuracy of combined detection of TuM2-PK ,CK18-3A9 and CYFRA21-1 were all better than TuM2-PK or CK18-3A9 alone(P<0 .05) .Con-clusion The combined detection of TuM2-PK ,CK18-3A9 and CYFRA21-1 were better in sensitivity and accuracy of diagnosis of NSCLC .

Noninvasive markers: a double-edged sword that stratifies nonalcoholic steatohepatitis

No abstract available.

Noninvasive predictors of nonalcoholic steatohepatitis in Korean patients with histologically proven nonalcoholic fatty liver disease

BACKGROUND/AIMS: The aims of this study were (1) to identify the useful clinical parameters of noninvasive approach for distinguishing nonalcoholic steatohepatitis (NASH) from nonalcoholic fatty liver disease (NAFLD), and (2) to determine whether the levels of the identified parameters are correlated with the severity of liver injury in patients with NASH. METHODS: One hundred and eight consecutive patients with biopsy-proven NAFLD (age, 39.8+/-13.5 years, mean+/-SD; males, 67.6%) were prospectively enrolled from 10 participating centers across Korea. RESULTS: According to the original criteria for NAFLD subtypes, 67 patients (62.0%) had NASH (defined as steatosis with hepatocellular ballooning and/or Mallory-Denk bodies or fibrosis > or =2). Among those with NAFLD subtype 3 or 4, none had an NAFLD histologic activity score (NAS) below 3 points, 40.3% had a score of 3 or 4 points, and 59.7% had a score >4 points. Fragmented cytokeratin-18 (CK-18) levels were positively correlated with NAS (r=0.401), as well as NAS components such as lobular inflammation (r=0.387) and ballooning (r=0.231). Fragmented CK-18 was also correlated with aspartate aminotransferase (r=0.609), alanine aminotransferase (r=0.588), serum ferritin (r=0.432), and the fibrosis stage (r=0.314). A fragmented CK-18 cutoff level of 235.5 U/L yielded sensitivity, specificity, and positive and negative predictive values of 69.0%, 64.9%, 75.5% (95% CI 62.4-85.1), and 57.1% (95% CI 42.2-70.9), respectively, for the diagnosis of NASH. CONCLUSIONS: Serum fragmented CK-18 levels can be used to distinguish between NASH and NAFL. Further evaluation is required to determine whether the combined measurement of serum CK-18 and ferritin levels improves the diagnostic performance of this distinction.

Apoptosis and diagnosis of nonalcoholic steatohepatitis / 대한간학회지

No abstract available.

Detection of chemosensitivity using K18-Asp(396) (M30) antibody in HeLa and OVCAR-3 cell lines treated with anticancer agents / 대한산부인과학회지

OBJECTIVE: The aim of this study was to detect the levels of M30-antigens as a biomarker of apoptosis in cells and their culture media after treatments with anticancer drugs as a preclinical study. METHODS: After HeLa and OVCAR-3 cells were treated respectively with paclitaxel, cisplatin, and camptothecin, the harvested cells were stained sequentially with M30 monoclonal antibodies and propidium iodide (PI). Afterwards, they were analyzed using a FACScan flow cytometer and observed under an immunofluorescence microscope for M30-FITC immunofluorescences. Levels of M30 antigens were also detected in their culture media using M30-Apoptosense ELISA kit. RESULTS: The levels of M30-FITC immunofluorescences were elevated in both cell lines after each drug treatments compared with those of control cells. The levels of M30 antigens detected by ELISA in media culturing each cell line treated with each of drugs were elevated compared with those of control cells. CONCLUSION: This study suggests that M30-antigens representing chemotherapy induced apoptosis may be a useful biomarker for predicting and monitoring the response of neoadjuvant chemotherapy in patients with gynecologic cancers.

Primary culture and hepatocelluar specific protein expression in human amniotic epithelial cells / 上海第二医科大学学报

Objective To optimize the method of primary culture for human amniotic epithelial cells (AECs) and detect the expression of hepatocelluar specific proteins in AECs. Methods AECs were obtained by collagenase and trypsin digestion method. 10,20,40 ng/mL epidermal growth factor (EGF) and 10 ng/mL basic fibroblast growth factor were added separately to the culture media,inverted microscope was used to observe the cell growth and proliferation,and the optimal condition for primary culture of AECs was obtained. The primary cells cultured without growth factors were served as controls. The expression of hepatocelluar specific proteins such as albumin,cytokeratin-18 (CK-18),alpha-1 antitrypsin (AAT) and alpha-1 foetoprotein (AFP) in cultured cells and histological sections were detected by immunohistochemical staining. Results The AECs available were relatively pure,with only a few mesenchymal cells. EGF of 10 ng/mL was chosen as an ingredient of culture medium as the growth and proliferation of AECs significantly accelerated with 10 ng/mL EGF. The expression of albumin,CK-18,AAT and AFP was detected in amnion tissues and AECs cultured in vitro. Conclusion Collagenase and trypsin digestion method and culture medium with 10 ng/mL EGF are favourable conditions for primary culture of AECs. Hepatocelluar specific proteins are expressed in human amnion tissues and AECs cultured in vitro,indicating that AECs have some characteristics of hepatocytes.