A novel diagnostic marker, p28(GANK) distinguishes hepatocellular carcinoma from potential mimics

PURPOSE. To investigate the sensitivity, specificity, and spatial distribution of the product of p28 gene (p28(GANK) protein) in human hepatocellular carcinoma (HCC) and nonhepatocellular carcinomas in relation to immunostaining with Cytokeratin 18 (CK18), alpha-fetoprotein (AFP), and Hepatocyte paraffin 1 (HepPar1). METHOD. In this retrospective study, formalin-fixed paraffin-embedded tissues from 24 HCCs, five intrahepatic cholangiocarcinomas (ICC), five combined hepatocellular cholangiocarcinomas (C-HCC-CC) and mine metastatic hepatic carcinomas (MHC) were immunostained for p28(GANK) as well as CK18, AFP and HepPar1. Only cases with more intensified staining in carcinoma contrast to the adjacent liver tissues were accepted as positive. RESULT. In HCC, p28(GANK) was expressed restrictively in hepatocytes of both para-lesion and carcinoma liver tissues, while absent in the bile duct epithelial cells, Kupffer cells, and other interstitial cells. The positive staining of p28(GANK) was noted in 16 (66.7％) specimens of HCC and three (60.0％) specimens of C-HCC-CC, and no specific lesion staining was found in all tested specimens of ICC and MHC. Sensitivity and specificity for hepatocyte-originated carcinoma were, respectively, 65.5％ and 100％ for p28(GANK), 79.3％ and 85.2％ for CK18, 20.7％ and 100％ for AFP, 79.3％ and 92.0％ for HepPar1. CONCLUSION. The hepatocytic staining for p28(GANK) is clearly useful in differentiating hepatocyte-originated carcinoma from non-HCC. p28(GANK) may be used as an ancillary marker for the diagnosis of HCC.     

Correlation and expression of COX-2 and P53 protein in basal cell carcinoma of eyelid

The correlation between the expression of COX-2 and p53 protein in basal cell carcinoma （BCC） of eyelid and apoptosis was investigated. Specimens of BCC were collected from 40 cases （aged 28-68 y） at the Department of Pathology, Renmin Hospital of Wuhan University, and Department of Pathology, Zhongnan Hospital of Wuhan University during from 1999 to 2006. Five specimens of paracancerous tissues served as control group. Immunohistochemical staining was performed to detect the expression of COX-2 and p53 in the tissues. The average absorbance （A） and the average positive area rate of COX-2 and p53 protein were measured by image analysis. The positive area rate of COX-2 and p53 protein was analyzed by linear correlation analysis. It was found that COX-2 and p53 proteins were highly expressed in BCC of eyelid, and weakly expressed in paracancerous tissues. Image analysis revealed that the expression of COX-2 and p53 proteins in BCC of eyelid was sig- nificantly higher than that in paracancerous tissues （P〈0.01）. Spearman rank correlation analysis demonstrated a positive correlation between the expression of COX-2 and p53 （r=0.113, P=0.421）. It was concluded that COX-2 can increase the expression of p53 protein, therefore suppressing apoptosis.     

Effects of tetrandrine on phenotypic modulation of vascular smooth muscle cells and expression of p38 MAPK as well as MKP-1 after intimal injury of rabbit carotid arteries

Objective： To study the effects of tetrandrine （Tet） on phenotypic modulation of vascular smooth muscle cells （VSMCs） and expression of p38 mitogen-activated protein kinase （p38MAPK） as well as mitogen-activated protein kinase phosphatase-1 （MKP-1） after vascular intimal injury. Methods： HE staining was used to analyze vascular morphology of sham-injured group, injured group and Tet-treated group at day 28. lmmunohistochemistry, Western blot and RT-PCR were respectively used to detect the expression change of smooth muscle a-actin （SMa-actin）, proliferation cell nuclear antigen （PCNA）, p38MAPK and MKP-1 of injured group and Tet group at days 7, 14 and 28 after balloon injury. Results： ① All layers of vascular wall in sham-injured group were intact at day 28. The neointimal area was significantly increased and the lumen area notably decreased in injured group at day 28. The neointimal proliferation in Tet treated group was less than that in injured group, and the lumen area of Tet group was significantly increased than that of injured group at day 28. ②Compared with the injured group, the expression of SMa-actin, PCNA, p38MAPK and MKP-1 of vascular wall in Tet group was no difference, and the neointimal proliferation condition was also basically as same as injured group at day 7 after injury. The expression of PCNA and p38MAKP in Tet group was obviously lower than that in injured group, and the expression of MKP-1 in Tet group was obviously higher than that in injured group at days 14 and 28 after injury. The expression of SMa-actin in Tet group was slightly higher than that in injured group at days 14 and 28 after injury. Conclusions： Tet could reduce neointimal proliferation by inhibiting VSMCs phenotypic modulation and p38MAPK signaling transduction pathway as well as its down regulation.     

Inhibitory effect of iron on in vitro proliferation of smooth muscle cells

Background Iron is a biocorrodible metal that might be used in bioabsorbable stents.This study investigated the effects at the cellular and protein levels of soluble divalent iron （ferrous gluconate） and soluble trivalent iron （ferric chloride） on the proliferation of human aortic smooth muscle cell （HASMC） in vitro.Methods The water-soluble tetrazolium （WST-1） test was used to evaluate the effect of iron on proliferation of HASMC and Western blotting was used to measure the levels of signaling proteins involved in proliferative and apoptosis pathways.Results HASMC proliferation was inhibited in a concentration dependent manner after treatment with soluble divalent and trivalent iron at concentrations of 100-500 μmol/L.Western blotting analysis showed that the proliferating cell nuclear antigen （PCNA） expression following treatment with soluble divalent iron and trivalent iron at 100,300 and 500 μmol/L was reduced compared to the control.The PCNA expression decreased with increasing iron concentration and to a greater extent with the trivalent iron than with the divalent iron treatment group.The p53 expression was markedly increased in a concentration dependent manner in both iron treatment groups.Conclusion The soluble divalent iron and,to a greater degree trivalent iron,inhibited HASMC proliferation in a dosedependent manner,which may be attributed to reduction of PCNA expression and increase of p53 expression.     

Expression of Survivin, p53 and its relationship with apoptosis, proliferation in hepatocellular carcinoma（HCC）

Objective:To investigate the expression of Survivin p53 and its relationship with apoptosis、proliferation in hepatocellular carcinoma(HCC). Methods:The expression of Survivin, p53 and the proliferation of tumor cells marked by proliferation cell nuclear antigen (PCNA) in 42 cases of HCC were assessed by immunohistochemical method. TUNEL was used to detect apoptosis. Results:Survivin protein was expressed in 30 of 42 cases of HCC(71.4%) and in 4 of 34 cases of adjacent cirrhosis tissues(11.8%). Expression of Survivin protein was negative in 10 cases of normal tissues. Survivin protein positive expression rate in HCC was significantly higher than adjacent cirrhosis tissues and normal tissues(P < 0.001). The increased Survivin protein expression in cancer was significantly associated with histological grade(P = 0.003), p53 protein(P = 0.013), survival time(P < 0.05) and the ratio of proliferative index to apoptotic index(P < 0.01), but was not significantly correlated with age, sex, clinical stage, tumor size, metastasis, AFP, HBs-Ag(P > 0.05).Conclusion:There is a marked increased expression of Survivin in HCC, which may play an important role in breaking the balance of proliferation and apoptosis of HCC cells. The correlation between Survivin and p53 expression in HCC indicates that cooperation between Survivin and p53 plays a certain role in occurrence and /or development of HCC.     

Lamin C protein deficiency in the primary fibroblasts from a new laminopathy case with ovarian cystadenoma

Background Laminopathies are a group of rare genetic disorders characterized by multiple-tissue degeneration.We describe a new laminopathy with ovarian cystadenoma and explore its molecular etiology.Methods The case is a 15-year-old girl who presents the prominent progeroid disorders, multiple system degeneration and early-onset cystadenoma of the ovary.Candidate genes including LMNA, ZMPSTE24, PPAR G, INSR and WRN were sequenced to screen for DNA variants.The mRNA and protein expression levels of LMNA were examined in primary fibroblasts.The pathophysiological events such as morphologic alterations, cell senescence, cell proliferation,apoptosis and pRb as well as p53 protein expressions were also investigated in primary fibroblasts.Results No mutation was identified in the candidate genes screened.Nuclear abnormalities including nuclear blebs,mislocalization of lamin A/C were evident in the patient fibroblasts.Ultrastructurally, nucleus exhibited nuclear herniation and almost complete loss of peripheral heterochromatin.In addition, lamin C protein expression was markedly reduced whereas lamin A protein level was normal and no prelamin A was detected in the primary fibroblasts.Although the senescence-associated β-galactosidase staining of patient＇ cells was negative, cells in S phase increased in accompany with a decrease in pRb protein expression.Furthermore, increases in apoptotic cell death and p53 expression were observed.Conclusions Our data suggest that selective deficiency of lamin C protein is associated with a case of laminopathy with ovarian cystadenoma.The abnormalities in nuclear structure and alterations in gene expression such as the decrease in pRb and increase in p53 may be responsible for the multiple tissue degeneration.     

Higher intratumor than peritumor expression of DUSP6/MKP-3 is associated with recurrence after curative resection of hepatocellular carcinoma

Background The MAPK phosphatases （MKPs） are a family of dual-specificity phosphatases （DUSPs） that can dephosphorylate both phosphothreonine and phosphotyrosine residues,thus inactivating MAPK signaling.DUSP6 is a cytoplasmic MKP that can inactivate ERK.DUSP6 has been implicated in the development of some tumors.The aim of this research was to investigate the expression of DUSP6 in hepatocellular carcinoma （HCC） and the correlation of DUSP6 with mitogen-activated protein kinases （MAPKs）,clinicopathological characteristics,and prognosis.Methods Tissues from 305 patients who had undergone hepatectomy for HCC was used in this study.The expression of DUSP6,p-ERK,p-JNK,and p-p38a was determined using tissue microarrays for immunohistochemical analysis.The prognostic value of DUSP6 and other clinicopathological factors were evaluated.Results The expression of DUSP6 was significantly higher in the tumor tissue when compared to the peritumor or normal liver tissue （P ＜0.001）.Tumor DUSP6 expression was significantly associated with disease-free survival （DFS） （P=0.013）.Tumor DUSP6 expression was an independent prognostic factor for DFS （Hazard ratio =1.635,P=0.006）.Conclusions DUSP6 is over expressed in tumor tissue compared to peritumor or normal liver tissue.Higher expression of DUSP6 in tumor tissue,than in peritumor tissue,is associated with the recurrence after curative resection of HCC,and the relative tumor DUSP6 expression has good power to predict the recurrence of HCC.