OBJECTIVETo investigate the relationship between SNC19 protein and cancer metastasis.

METHODSExpression of SNC19 protein in cancer cell lines and tissues was examined by Western blot analysis using anti-SNC19 monoclonal antibody. In addition, Psectag2A-SNC19(ORF) eukaryotic expression vector was constructed and transfected into BCAP37 cells. After the target protein was expressed and purified, processing forms of SNC19 protein were further identified using anti-His mAb and each form was assayed for its gelatinase activity.

RESULTSDifferent expression and post-translational processing of the SNC19 proteins in the cancer cell lines and intestinal tissues were detected.BCAP37 cells transfected full-length SNC19 (ORF) generated two different sized proteins in cell lysates, 120 and 75 kD; 75 kD was detected to have proteolytic activity by gelatin zymography.

CONCLUSIONSNC19 protein presents different expression and post-translational processing in the cancer cells and tissues, of which 75 kD was identified to have gelatinase activity.

Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; DNA, Complementary ; chemistry ; Female ; Gelatinases ; metabolism ; Humans ; Neoplasm Metastasis ; Neoplasm Proteins ; chemistry ; genetics ; metabolism ; Protein Processing, Post-Translational ; Serine Endopeptidases ; chemistry ; genetics ; metabolism ; Transfection

Animals ; Annexin A2 ; chemistry ; genetics ; metabolism ; physiology ; DNA Replication ; Endocytosis ; Exocytosis ; Humans ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasms ; metabolism ; pathology ; Prognosis ; Tissue Plasminogen Activator ; metabolism ; physiology

OBJECTIVETo screen the differentially methylated DNA sequences between mucosa adjacent to colorectal cancer (MACC) and normal colonic mucosa.

METHODSThe methylated DNA sequences were enriched by methylation CpG islands amplification (MCA), and the differentially methylated DNA sequences between MACC and normal colonic mucosa were isolated by representational difference analysis (RDA). Similarities between the separated fragments and the human genomic DNA were analyzed with BLAST program system in GenBank. With the separated fragment 1A12 as probe, dot blot was used to study its distribution between RDA products (No. 1-4 rounds), MACC (tester) and normal colonic mucosa(driver).

RESULTSTwenty-five differentially methylated DNA sequences were obtained. Preliminary studies indicated that 1A01 fragment was concerned with two different genes (LOC256866 and CECR7), it was located in the first exon of CECR7. 1A12 fragment was located in 5 flanking region of GR6 gene. By dot blot with 1A12 probe, hybridized signals were detected in MCA product of MACC and RDA products of No. 1-4 rounds, respectively. No signal was detected in MCA product of normal colonic mucosa.

CONCLUSIONThe differentially methylated DNA sequences can be isolated effectively between two different tissues with MCA coupled with RDA. Different methylated DNA fragments exist between MACC and normal colonic mucosa and these fragments may be concerned with colorectal cancer.

Adult ; Base Sequence ; Colorectal Neoplasms ; diagnosis ; genetics ; CpG Islands ; genetics ; DNA Methylation ; DNA, Neoplasm ; chemistry ; isolation & purification ; metabolism ; Genetic Testing ; Humans ; Intestinal Mucosa ; metabolism ; Polymerase Chain Reaction ; Sequence Analysis, DNA

To establish a quantitative assay for telomerase activity and analyze the telomerase activity in peripheral blood mononuclear cells (PBMNC) from patients with acute leukemia, a fluorescent dye, PicoGreen, was added to the products after telomere repeat amplification protocol. The samples were excited at 480 nm and the fluorescence emission intensity was measured at 520 nm using a spectrofluorometer. Telomerase activity was detected in PBMNCs from 20 cases of normal individuals and 25 patients with acute leukemia. The results showed that the fluorescence of PicoGreen binding to double-stranded DNA specifically was enhanced with increase of DNA quantities. In conclusion, the met hod is rapid, simple and quantitative, the telomerase activities of PBMNCs from acute leukemia patients are significantly higher than that of the normal controls.
Acute Disease ; Adolescent ; Adult ; Aged ; Cell Line ; DNA, Neoplasm ; genetics ; metabolism ; Female ; Fluorescent Dyes ; chemistry ; Humans ; Leukemia ; blood ; enzymology ; genetics ; Leukocytes, Mononuclear ; enzymology ; Male ; Middle Aged ; Organic Chemicals ; Telomerase ; chemistry ; genetics ; metabolism

Coexpression of Kit ligand and c-kit has been reported in some gynecologic tumors. To determine whether imatinib mesylate is useful in ovarian epithelial tumors, we performed immunohistochemical and mutational analysis. The cases consisted of 33 cases, which included 13 serous cystadenocarcinomas, 1 borderline serous tumor, 8 mucinous cystadenocarcinomas, 6 borderline mucinous tumors and 5 clear cell carcinomas. Five cases of serous cystadenoma and 5 cases of mucinous cystadenoma were also included. In the immunohistochemical study, 3 cases (3/6, 50%) of borderline mucinous cystic tumor and two cases (2/8, 25%) of mucinous cystadenocarcinoma show positive staining for KIT protein. Only one case (1/13, 7.7%) of serous cystadenocarcinoma had positive staining. On mutational analysis, no mutation was identified at exon 11. However, two cases of borderline mucinous tumors and one case of mucinous cystadenocarcinoma had mutations at exon 17. In these cases, the immunohistochemistry also shows focal positive staining at epithelial component. Although, KIT protein expression showed higher incidence in mucinous tumors than serous tumors, they lack KIT-activating mutations in exon 11. Thus, ovarian surface epithelial tumors are unlikely to respond to imatinib mesylate.
Adult ; Aged ; Cystadenocarcinoma, Mucinous/genetics/metabolism/pathology ; Cystadenoma, Mucinous/genetics/metabolism/pathology ; Cystadenoma, Serous/genetics/metabolism/pathology ; DNA Mutational Analysis ; DNA, Neoplasm/chemistry/genetics ; Epithelial Cells/chemistry/metabolism/pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Middle Aged ; Mutation ; Ovarian Neoplasms/genetics/metabolism/*pathology ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Proto-Oncogene Proteins c-kit/biosynthesis/*genetics

OBJECTIVETo observe the effect on P170, LRP, TOPO II of S180 tumour MDR mice for matter by 70% ethanol with Huanglian Jiedu Tang, and then discuss the molecular biology base for clinic.

METHOD18-22 gramme mice were divided into four groups for normal S180 tumour cell group, matter by 70% ethanol with Huanglian Jiede Tang 100 mg x kg(-1) and 50 mg x kg(-1) in random. Each mouse was given S180 cell 0.2 mL by celiac, and after 24 hours give cisplatin for Injective 3 mg x kg(-1), ip, once a week. And give cyclophosphamide and 5-FU 3 mg x kg(-1), ig, once every day. After 15 days, collect lively mice ascites and give it for onefold normal mice. And then repeat before process. At the same time, every group was given corresponding medicine for 0.2 mL x 10 g(-1). The normal group and the model group were given the same cubage water, all together fore weeks. At last observd the P170, LRP, TOPO II by flow cytometry.

RESULTMatter by 70% ethanol with Huanglian Jiedu Tang could obviously reduce the express of P170 and LRP, and the activiation of TOPO II.

CONCLUSIONMatter by 70% ethanol with Huanglian Jiedu Tang can intervene the ocurrence of the multi-drug resistance of tumour cells by regulating the biology gene.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Cell Line, Tumor ; Coptis ; chemistry ; DNA Topoisomerases, Type II ; metabolism ; Drug Combinations ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Ethanol ; chemistry ; Flow Cytometry ; Mice ; Phytotherapy ; Plants, Medicinal ; chemistry ; Sarcoma 180 ; metabolism ; pathology ; prevention & control ; Vault Ribonucleoprotein Particles ; metabolism

This paper gave a brief introduction of the effect of Solanum nigrum on anti-cancer. The experimental results showed that the total alkaloid isolated from S. nigrum interfered structure and function of tumor cell membrane, disturbed the synthesis of DNA and RNA, changed the cell cycle distribution, so that total alkaloids could play in inhibabition to tumor cells, while the glycoprotein (150 x 10(3)) isolated from S. nigrum might have shown anti-cancer abilities by blocking the anti-apoptotic pathway of NF-kappaB, activating caspase cascades reaction and increasing the production of nitric oxide.
Alkaloids ; isolation & purification ; pharmacology ; Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Membrane Permeability ; drug effects ; DNA, Neoplasm ; biosynthesis ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Glycoproteins ; isolation & purification ; pharmacology ; Humans ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; RNA, Neoplasm ; biosynthesis ; Sialic Acids ; metabolism ; Solanum nigrum ; chemistry

OBJECTIVETo clone human gastric cancer related gene and to analyze its expression profile in gastric mucosal tissues.

METHODSPaired tumor, paratumor and non-tumor specimens from 7 gastric adenocarcinoma patients (male 4, female 3, with average age 51 +/- 18 years) were studied by means of fluorescent differential display reverse transcription polymerase chain reaction (DDRT-PCR). The differentially expressed cDNA bands of interest were cloned and analyzed by Northern blot and in situ hybridization. Thirty cases (male 23 female 7 with average age 59 +/- 8 years) of paired paraffin-embedded gastric tumor and non-tumor tissues were used in in situ hybridization analysis.

RESULTSA gene expressed much lower in 6 out of 7 tested tumor samples than in their normal and paratumor counterparts was identified. It was named GCRG123. Northern blot analysis confirmed the differential expression. Human multiple tissue Northern blot analysis showed that GCRG123 expressed in various adult human tissues including thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocyte. Sequence analysis revealed that GCRG123 (GenBank accession number AF454554) was a lamin like protein gene. It had one open reading frame which consisted of 49 amino acids (GenBank accession number AAL61668.1). In situ hybridization analysis showed a high GCRG123 expression level in normal gastric epithelium and pylori glands, but low expression level in tumor as well as dysplasia and most intestinal metaplasia at the paratumor regions.

CONCLUSIONA lamin-like protein gene was identified in human gastric mucosa, it is down-regulated in gastric cancer and its precancerous leisions.

Adult ; Aged ; Blotting, Northern ; Cloning, Molecular ; DNA, Neoplasm ; chemistry ; genetics ; Down-Regulation ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; In Situ Hybridization ; Lamins ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; RNA, Messenger ; genetics ; metabolism ; Sequence Analysis, DNA ; Stomach Neoplasms ; genetics

The article submitted the new progress of the study in antitumor mechanism of bererine (Ber). Reports indicated that Ber suppressed growth of tumor cells through impacting tumor cells growth cycle, inhibiting synthesises of DNA and protein, and reducing the activity of topoisomerase. Ber improved tumor cells apoptosis through several ways such as regulating apoptotic gene expression, inducing the decline of transmembrane potential on mitochondria. And Ber still could inhibit tumor metastasis through suppressing the formation of tumor angiogenesis, blocking signal transduction pathway, antagonizing extralumen, et al. In addition, Ber could induce tumor cells to differentiate to antagonize tumor.
Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Berberine ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA, Neoplasm ; biosynthesis ; Humans ; Neoplasm Proteins ; biosynthesis ; Neoplasms ; genetics ; metabolism ; pathology ; Plants, Medicinal ; chemistry

OBJECTIVETo study the alterations of FHIT gene and P16 gene in malignant transformed human bronchial epithelial cells induced by crystalline nickel sulfide using an immortal human bronchial epithelial cell line, and to explore the molecular mechanism of nickel carcinogenesis.

METHODS16HBE cells were treated 6 times with different concentrations of NiS in vitro, and the degree of malignant transformation was determined by assaying the anchorage-independent growth and tumorigenicity. Malignant transformed cells and tumorigenic cells were examined for alterations of FHIT gene and P16 gene using RT-PCR, DNA sequencing, silver staining PCR-SSCP and Western blotting.

RESULTSNiS-treated cells exhibited overlapping growth. Compared with that of negative control cells, soft agar colony formation efficiency of NiS-treated cells showed significant increases (P < 0.01) and dose-dependent effects. NiS-treated cells could form tumors in nude mice, and a squamous cell carcinoma was confirmed by histopathological examination. No mutation of exon 2 and exons 2-3, no abnormal expression in p16 gene and mutation of FHIT exons 5-8 and exons 1-4 or exons 5-9 were observed in transformed cells and tumorigenic cells. However, aberrant transcripts or loss of expression of the FHIT gene and Fhit protein was observed in transformed cells and tumorigenic cells. One of the aberrant transcripts in the FHIT gene was confirmed to have a deletion of exon 6, exon 7, exon 8, and an insertion of a 36 bp sequence replacing exon 6-8.

CONCLUSIONSThe FHIT gene rather than the P16 gene, plays a definite role in nickel carcinogenesis. Alterations of the FHIT gene induced by crystalline NiS may be a molecular event associated with carcinogen, chromosome fragile site instability and cell malignant transformation. FHIT may be an important target gene activated by nickel and other exotic carcinogens.

Acid Anhydride Hydrolases ; chemistry ; genetics ; metabolism ; Animals ; Base Sequence ; Bronchi ; cytology ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; DNA Damage ; Exons ; Gene Deletion ; Genes, p16 ; Humans ; Mice ; Mice, Nude ; Molecular Sequence Data ; Mutagenicity Tests ; Neoplasm Proteins ; chemistry ; genetics ; metabolism ; Nickel ; toxicity ; RNA, Messenger ; metabolism ; Respiratory Mucosa ; cytology ; drug effects ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA