Expression of cytokeratin(CK)7, CK8/18, CK19 and p40 in esophageal squamous cell carcinoma and their correlation with prognosis / 中华病理学杂志

Objective@#To evaluate the expression of cytokeratin (CK)7, CK8/18, CK19 and p40 in esophageal squamous cell carcinoma (ESCC) and its significances. @*Methods@#One hundred and ninety cases of surgically resected ESCCs and 154 normal esophageal tissues as control were collected at the First Affiliated Hospital of Zhengzhou University in 2012.Of the 190 ESCC cases including 116 male and 74 female, aged 28-82 (60.3±8.6) years, 88 cases <60 years old and 102 cases ≥60 years old. Tissue sections were immunostained for CK7, CK8/18, CK19 and p40, and the expression was evaluated and correlated with the clinicopathologic findings and outcome. @*Results@#CK19 and p40 were expressed in 190 cases of ESCCs; with 147 cases (77.4%) and 151 cases (79.5%) showing high p40 and CK19 expression, respectively; while 43 cases (22.6%) and 39 cases (20.5%) showed low p40 and CK19 expression, respectively. The low expression groups showed more lymph node metastases and higher pTNM stages compared to the high expression groups. The high CK19 expression group showed better prognosis than the low expression group (P<0.01); p40 expression was not correlated with prognosis(P>0.05). In contrast, CK7 and CK8/18 expression was only seen in 29 cases (15.3%) and 59 cases (31.1%) of ESCCs, respectively, and their expression correlated significantly with the degree of tumor differentiation and lymph node metastasis (P<0.05). The prognosis in the CK7 negative group was better than that in the CK7 positive group. Similar results were found in CK8/18 expression. Multivariate analysis revealed that pTNM stages, low CK19 expression and CK8/18 expression were independent prognostic factors. @*Conclusions@#Low p40 expression and the expression of CK7 and CK8/18 cannot exclude poorly-differentiated ESCCs.CK7 and CK8/18 expression and low CK19 and p40 expression in the ESCCs are associated with lymph node metastasis and poor prognosis. Decreased expression of CK19 and positive expression of CK8/18 in ESCCs are independent prognostic markers.

The influence of intracellular keratin 8 on hepatitis C virus replication / 药学学报

The level of intracellular keratin 8(KRT-8) is associated with liver diseases, whose expression is increased in hepatitis C virus (HCV)-infected patients with hepatocarcinoma and in cultural cells infected with HCV. However, it is not clear whether KRT-8 will impact HCV replication. In this paper, the HCV replication was analyzed in response to high expression and silence of KRT-8. The inhibitory activities against wild-type and mutant HCV were also analyzed by silence of KRT-8 or combined with known anti-HCV drug telaprevir. Results showed that the protein level of KRT-8 was increased in proportion with the HCV replication. The high expression was found to facilitate HCV replication, while the silence of KRT-8 was able to inhibit HCV replication and enhanced the anti-HCV activity of telaprevir. It also inhibited A156T and D168V mutant HCV, which are resistant to protease inhibitors. These results suggest that KRT-8 is a co-factor for HCV replication. Down-regulation of KRT-8 can inhibit wild type and mutant HCV replication to enhance the anti-HCV activity of known anti-HCV drugs. Therefore, KRT-8 may be a new target in the development of anti-HCV agents.

12-O-Tetradecanoylphorbol-13-Acetate Induces Keratin 8 Phosphorylation and Reorganization via Expression of Transglutaminase-2

The stiffness of cancer cells is attributable to intermediate filaments such as keratin. Perinuclear reorganization via phosphorylation of specific serine residue in keratin is implicated in the deformability of metastatic cancer cells including the human pancreatic carcinoma cell line (PANC-1). 12-O-Tetradecanoylphorbol-13-acetate (TPA) is a potent tumor promoter and protein kinase C (PKC) activator. However, its effects on phosphorylation and reorganization of keratin 8 (K8) are not well known. Therefore, we examined the underlying mechanism and effect of TPA on K8 phosphorylation and reorganization. TPA induced phosphorylation and reorganization of K8 and transglutaminase-2 (Tgase-2) expression in a time- and dose-dependent manner in PANC-1 cells. These effects peaked after 45 min and 100 nM of TPA treatment. We next investigated, using cystamine (CTM), Tgase inhibitor, and Tgase-2 gene silencing, Tgase-2's possible involvement in TPA-induced K8 phosphorylation and reorganization. We found that Tgase-2 gene silencing inhibited K8 phosphorylation and reorganization in PANC-1 cells. Tgase-2 gene silencing, we additionally discovered, suppressed TPA-induced migration of PANC-1 cells and Tgase-2 overexpression induced migration of PANC-1 cells. Overall, these results suggested that TPA induced K8 phosphorylation and reorganization via Tgase-2 expression in PANC-1 cells.

Ethacrynic Acid Inhibits Sphingosylphosphorylcholine-Induced Keratin 8 Phosphorylation and Reorganization via Transglutaminase-2 Inhibition

Sphingosylphosphorylcholine (SPC) is significantly increased in the malicious ascites of tumor patients and induces perinuclear reorganization of keratin 8 (K8) filaments in PANC-1 cells. The reorganization contributes to the viscoelasticity of metastatic cancer cells resulting in increased migration. Recently, we reported that transglutaminase-2 (Tgase-2) is involved in SPC-induced K8 phosphorylation and reorganization. However, effects of Tgase-2 inhibitors on SPC-induced K8 phosphorylation and reorganization were not clearly studied. We found that ethacrynic acid (ECA) concentration-dependently inhibited Tgase-2. Therefore, we examined the effects of ECA on SPC-induced K8 phosphorylation and reorganization. ECA concentration-dependently suppressed the SPC-induced phosphorylation and perinuclear reorganization of K8. ECA also suppressed the SPC-induced migration and invasion. SPC induced JNK activation through Tgase-2 expression and ECA suppressed the activation and expression of JNK in PANC-1 cells. These results suggested that ECA might be useful to control Tgase-2 dependent metastasis of cancer cells such as pancreatic cancer and lung cancers.

Effect of HCMV on expressions of K8 and K18 in duct epithelial cells of salivary gland / 中华微生物学和免疫学杂志

Objective To study the effect of human cytomegalovirus (HCMV) on expressions of K8 and K18 in duct epithelial cells of salivary gland. Methods The expressions of immediate early antigen of HCMV, K8 and K18 were detected by immunohistochemistry staining in tissues embedded in paraffin of parotid cytomegalic inclusion disease(PCID). Results Cytomegly bearing inclusion appeared in duct epithelium of PCID. DDG9/CCH2 antigen of HCMV was expressed in cytomegly bearing inclusion. K8 was negative in these cytomegly while K18 was intensively positive. Conclusion It is suggested that breaking down of K8 be induced in parotid duct epithelial cells infected by HCMV and that up-regulation of K18 may be a reactive change. Keratin network in simple epithelium functions to impart mechanical integrity to cells.