[Antisense to CDK4 inhibits the growth of human colon cancer cells HT29].

OBJECTIVE: Antisense CDK4 was transfected into human colorectal cancer cells HT29 to examine the effect of it on the growth and proliferation of HT29 cells. METHODS: Antisense CDK4 transfected the HT29 cells with lipofectamine. The transfected effects were verified by the Northern blot, Western blot, morphological method, flow cell measure, etc. RESULTS: The transformant cells showed the expression of exogenous antisense CDK4 mRNA, and downmodulation of endogenous CDK4 mRNA expression and inhibition of its protein synthesis. The transformant cells exhibited the partial reversion of the malignant phenotype and behavior, and that the cell growth and the ability of colony formation in soft agar were inhibited. The arrest of G1 phase was also revealed. the apoptosis rate is higher in HT29-asCDK4. CONCLUSION: The antisense CDK4 gene can inhibited the cell growth and proliferation, and induce the cell apoptosis. It might provide a new way to gene therapy of the colorectal carcinoma.

Increased expression of mitogen-activated protein kinase and its upstream regulating signal in human gastric cancer.

AIM: To investigate the expression of mitogen-activated protein kinases (MAPKs) and its upstream protein kinase in human gastric cancer and to evaluate the relationship between protein levels and clinicopathological parameters. METHODS: Western blot was used to measure the expression of extracellular signal-regulated kinase (ERK)-1, ERK-2, ERK-3, p38 and mitogen or ERK activated protein kinaseMEK-1 proteins in surgically resected gastric carcinoma, adjacent normal mucosa and metastatic lymph nodes from 42 patients. Immunohistochemistry was employed for their localization. RESULTS: Compared with normal tissues, the protein levels of ERK-1 (integral optical density value 159 526+/-65 760 vs 122 807+/-65 515, P = 0.001), ERK-2 (168 471+/-95 051 vs 120 469+/-72 874, P<0.001), ERK-3 (118 651+/-71 513 vs 70 934+/-68 058, P<0.001), P38 (104 776+/-51 650 vs 82 930+/-40 392, P = 0.048) and MEK-1 (116 486+/-45 725 vs 101 434+/-49 387, P = 0.027) were increased in gastric cancer tissues. Overexpression of ERK-3 was correlated to TNM staging (average ratio of integral optic density (IOD)(tumor): IOD(normal) in TNM I, II, III, IV tumors was 1.43+/-0.34, 5.08+/-3.74, 4.99+/-1.08, 1.44+/-1.02, n = 42, P = 0.023) and serosa invasion (4.31+/-4.34 vs 2.00+/-2.03, P = 0.037). In poorly differentiated cancers (n = 33), the protein levels of ERK-1 and ERK-2 in stage III and IV tumors were higher than those in stage I and II tumors (2.64+/-3.01 vs 1.01+/-0.33, P = 0.022; 2.05+/-1.54 vs 1.24+/-0.40, P = 0.030). Gastric cancer tissues with either lymph node involvement (2.49+/-2.91 vs 1.03+/-0.36, P = 0.023; 1.98+/-1.49 vs 1.24+/-0.44, P = 0.036) or serosa invasion (2.39+/-2.82 vs 1.01+/-0.35, P = 0.022; 1.95+/-1.44 vs 1.14+/-0.36, P = 0.015) expressed higher protein levels of ERK-1 and ERK-2. In Borrmann II tumors, expression of ERK-2 and ERK-3 was increased compared with Borrmann III tumors (2.57+/-1.86 vs 1.23+/-0.60, P = 0.022; 5.50+/-5.05 vs 1.83+/-1.21, P = 0.014). Borrmann IV tumors expressed higher p38 protein levels. No statistically significant difference in expression of MAPKs was found when stratified to tumor size or histological grade (P>0.05). Protein levels of ERK-2, ERK-3 and MEK-1 in metastatic lymph nodes were 2-7 folds higher than those in adjacent normal mucosa. The immunohistochemistry demonstrated that ERK-1, ERK-2, ERK-3, p38 and MEK-1 proteins were mainly localized in cytoplasm. The expression of MEK-1 in gastric cancer cells metastasized to lymph nodes was higher than that of the primary site. CONCLUSION: MAPKs, particularly ERK subclass are overexpressed in the majority of gastric cancers. Overexpression of ERKs is correlated to TNM staging, serosa invasion, and lymph node involvement. The overexpression of p38 most likely plays a prominent role in certain morphological subtypes of gastric cancers. MEK-1 is also overexpressed in gastric cancer, particularly in metastatic lymph nodes. Upregulation of MAPK signal transduction pathways may play an important role in tumorigenesis and metastatic potential of gastric cancer.

[The retrospective analysis of HBV and HCV infection in cholangiocarcinoma].

OBJECTIVE: In order to study the diagnosis and treatment of HBV and HCV infection. METHODS: We retrospectively analysed clinical data of 680 patients with cholangiocarcinoma from 1995 to 2001 and stated by SPSS software. RESULTS: (1) The fastigium of cholangiocarcinoma was 60 - 65 years old. The incidence of cholangiocarcinoma was higher in aged males and the sex ratio (male:female) was 1.36:1. (2) The proximal cholangiocarcinoma was most (41.6%) and distant cholangiocarcinoma was secondly (28.7%). (3) Most patients of cholangiocarcinoma were late. The resection rate was low and the rate of radical operation was 21.6% (147/680). (4) The incidence of proximal cholangiocarcinoma was higher in the positive Serologic marks for HBV and HCV and course of diseases was short. Moreover, the pathology of. positive Serologic marks for HBV and HCV trended to low-differentiation and invasion, metastasis and the resection rate was lower. CONCLUSIONS: Cholangiocarcinoma is common in the aged males. The infection of HB(C)V and hilar cholangiocarcinoma are correlated and incline to the proximal bile duct. The hilar cholangiocarcinoma infected HB(C)V may have higher malignant degree in biological characteristics and more badly prognosis.

Cholangiocarcinoma cells express somatostatin receptor subtype 2 and respond to octreotide treatment.

BACKGROUND/PURPOSE: We investigated the in vitro and in vivo inhibitory effects of a somatostatin analogue (octreotide, OCT) on cholangiocarcinoma cell lines. METHODS: The reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to detect the gene expression of five somatostatin receptor (SSTR) subtypes in four cholangiocarcinoma cell lines (RBE, NEC, QBC939, and SSP-25). The antiproliferative effects of OCT on these cell lines were determined by means of an MTT assay in vitro, as well as in a nude mouse tumor heterograft model in vivo. Apoptosis and cell cycles in the cholangiocarcinoma cell lines after OCT administration were evaluated by flow cytometry; and the effects of OCT on the expression of cyclin E, cyclin-dependent kinase 2 (CDK2), and p27kipl were evaluated by Western blots. RESULTS: Only SSTR2 mRNA was detected in these four cholangiocarcinoma cell lines. OCT significantly inhibited the proliferation of the four cholangiocarcinoma cell lines in vitro ( P < 0.05 vs control), and the weights of the QBC939 xenografts in the OCT-treated group were lower than those in the control group, but there was no significant difference between them. After 48-h exposure to 10(3) ng/ml OCT, flow cytometric analysis demonstrated an increased number of cells in G0/G1 phase associated with a decreased number of cells in G2/M and S phases ( P < 0.01 vs control). Apoptosis was not observed in any samples. The expression of p27kipl was promoted by OCT administration, while that of cyclin E and that of CDK2 were inhibited. CONCLUSIONS: The results proved that OCT inhibits the proliferation of cholangiocarcinoma cells through G0/G1 cell cycle arrest rather than through the process of apoptosis. These effects are partially mediated by enhancing the expression of p27kipl, and decreasing the amounts of cyclin E-CDK2 complex.