Gene expression profiles of hepatoma cell line HLE

AIM: To investigate the global gene expression of cancer related genes in hepatoma cell line HLE using Atlas Human Cancer Array membranes with 588 well-characterized human genes related with cancer and tumor biology.METHODS: Hybridization of cDNA blotting membrane was performed with 32P-labeled cDNA probes synthesized from RNA isolated from Human hepatoma cell line HLE and noncirrhotic normal liver which was liver transplantation donor.AtlasImage, a software specific to array, was used to analyze the result. The expression pattern of some genes identified by Atlas arrays hybridization was confirmed by reverse transcription polymerase chain reaction (RT-PCR)in 24 pairs of specimens and Northern blot of 4 pairs of specimens.RESULTS: The differential expression of cell cycle/growth regulator in hepatocellular carcinoma (HCC) showed a stronger tendency toward cell proliferation with more than 1.5-fold up-regulation of Cyclin C, ERK5, ERK6, E2F-3, TFDP2 and CK4. The anti-apoptotic factors such as Akt-1 were up-regulated, whereas the promotive genes of apoptosis such as ABL2 were down-regulated. Among oncogene/tumors suppressors, SKY was down-regulated. Some genes such as Integrin beta 8, Integrin beta 7, DNA-PK, CSPCP,byglycan, Tenacin and DNA Topo were up-regulated. A number of genes, including LAR, MEK1, eps15, TDGF1,ARHGDIA were down-regulated. In general, expression of the cancer progression genes was up-regulated, while expression of anti-cancer progression genes was downregulated. These differentially expressed genes tested with RT-PCR were in consistent with cDNA array findings.CONCLUSION: Investigation of these genes in HCC is helpful in disclosing molecular mechanism of pathogenesis and progression of HCC. For the first time few genes were discovered in HCC. Further study is required for the precise relationship between the altered genes and their correlation with the pathogenesis of HCC.

K-ras gene mutation in the diagnosis of ultrasound guided fine-needle biopsy of pancreatic masses

AIM: To investigate the utility of K-ras mutation analysis ofultrasound guided fine-needle aspirate biopsy of pancreaticmasses.METHODS: Sixty-six ultrasound guided fine-needle biopsieswere evaluated by cytology, histology and k-ras mutation.The mutation at codon 12 of the k-ras oncogene wasdetected by artificial restriction fragment lengthpolymorphisms using Bst NI approach.RESULTS: The presence of malignant cells was reported in40 of 54 pancreatic carcinomas and K-ras mutations weredetected in 45 of the 54 FNABs of pancreatic carcinomas. Thesensitivity of cytology and k-ras mutation were 74 % and 83%, respectively. The speciality of cytology and k-ras mutationwere both 100 %. The sensitivity and speciality of k-ras mutationcombined with cytology were 83 % and 100 %, respectively.CONCLUSION: High diagnostic accuracy with acceptablediscomfort of FNAB make it useful in diagnosis of pancreaticcarcinoma. Ultrasound guided fine-needle biopsy is a safeand feasible method for diagnosing pancreatic cancer.Pancreatic carcinoma has the highest K-ras mutation rateamong all solid tumors. The mutation rate of k-ras is about80-100 %. The usage of mutation of codon 12 of k-rasoncogene combined with cytology is a good alternative forevaluation of pancreatic masses.