OBJECTIVETo investigate the effect of fragile histidine triad (FHIT) gene on the apoptosis of gastric cancer cells.

METHODSFHIT gene was transfected into gastric cancer cell line MGC-803 by liposome. Western blot and immunohistochemical assay were employed to detect the expression of FHIT. Apoptosis and cell cycle were analyzed by flow cytometry (FCM). Morphological changes were observed by light and electron microscope.

RESULTSStable expression of FHIT protein in transfected MGC-803 cells was obtained. The rates of G0/G1 phase and apoptosis cells were significantly higher in FHIT-transfected MGC-803 cells than those in control cells (64.4%compared with 49.5%and 32.0%compared with 12.2%, respectively).

CONCLUSIONThe results indicate that FHIT gene might be involved in the apoptosis of gastric cancer cells.

Acid Anhydride Hydrolases ; Apoptosis ; Cell Line, Tumor ; Flow Cytometry ; Genes, Tumor Suppressor ; Humans ; Immunohistochemistry ; Neoplasm Proteins ; analysis ; genetics ; Stomach Neoplasms ; genetics ; pathology ; prevention & control ; Transfection

OBJECTIVETo evaluate the effect of a novel blockade technique for gastric cancer on blood-borne metastasis of gastric cancer cells to portal vein.

METHODSTwenty-three cases of gastric cancer were divided into routine operation group (8 cases intraoperatively without blockade technique) and blockade group (15 cases with blockade technique). Blood samples from portal vein pre- and intraoperatively, as well as gastroepiploic vein limited within the blockade area were obtained to detect CK19 mRNA expression by using RT-PCR technique.

RESULTSBefore the dissection of gastric lesion, the overall positive rate of CK19 mRNA expression in portal vein blood is 34.7% (9/23), including 37.5% (3/8) in routine operation group and 33.3% (5/15) in blockade group. While the course of tumor resection, those positive rates were 87.5% (7/8) in routine operation group and 6.7% (1/15) in blockade group respectively (P < 0.05). CK19 mRNA expression in the right gastroepiploic venous blood limited within the blocking area was all positive in 15 cases of blockade group.

CONCLUSIONThis blockade technique can be used effectively to block the intraoperative spread of gastric cancer cells, thus prevent blood-borne metastasis due to operative manipulation.

Aged ; Biomarkers, Tumor ; blood ; genetics ; Female ; Gastrectomy ; Humans ; Keratins ; blood ; genetics ; Ligation ; Male ; Middle Aged ; Neoplasm Metastasis ; prevention & control ; Neoplastic Cells, Circulating ; pathology ; RNA, Messenger ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; blood ; pathology ; surgery ; Vascular Surgical Procedures ; methods

OBJECTIVESTo study the effects of c9,t11-conjugated linoleic acid (c9,t11-CLA) on invasive ability of human gastric carcinoma cell line (SGC-7901) and to explore its possible mechanism.

METHODSReconstituted basement membrane invasion assay was used to evaluate invasive ability of cancer cells. Expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA was measured by reverse transcription polymerase chain reaction (RT-PCR) in SGC-7901 cells.

RESULTSAt the concentrations of 200 micromol/L, 100 micromol/L and 50 micromol/L, c9,t11-CLA suppressed their reconstituted basement membrane invasion of SGC-7901 by 53.7%, 40.9% and 29.3%, respectively. c9,t11-CLA could induce the expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA in SGC-7901 cells.

CONCLUSIONSThe invasion of SGC-7901 cells could be inhibited by c9,t11-CLA through reconstituted basement membrane. Anti-invasion action of c9,t11-CLA might be associated with induction of expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA in tumor cells.

Adenocarcinoma ; pathology ; Gene Expression ; drug effects ; Humans ; Linoleic Acid ; pharmacology ; therapeutic use ; Monomeric GTP-Binding Proteins ; biosynthesis ; genetics ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Invasiveness ; prevention & control ; Nucleoside-Diphosphate Kinase ; RNA, Messenger ; biosynthesis ; drug effects ; Stomach Neoplasms ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics ; Transcription Factors ; biosynthesis ; genetics ; Tumor Cells, Cultured

The E2F-1 transcription factor is post-translationally modified and stabilized in response to various forms of DNA damage to regulate the expression of cell-cycle and pro-apoptotic genes. The sustained overexpression of E2F-1 is a characteristic feature of gastric cancer. In this study, we investigated the role of short hairpin RNA (shRNA) targeting E2F-1 gene on human gastric cancer MGC-803 cell growth in vivo, and preliminarily revealed the mechanism. Thus, we constructed recombinant pGCSIL-GFP-shRNA-E2F-1 lentiviral vector to knock down E2F-1 expression in human gastric cancer MGC-803 cells in vivo, and studied the effect of E2F-1 shRNA on growth of MGC-803 tumor and evaluated its treatment efficacy. Our data demonstrated that in a mouse model of established gastric cancer, intratumor injection of lentiviral shRNA targeting E2F-1 definitely decreased the endogenous E2F-1 mRNA and protein expression in MGC-803 tumor, and inhibited tumor growth and promoted tumor cells apoptosis. Moreover, we found that E2F-1 shRNA increased the expression of phosphatase and tensin homolog (PTEN), activated caspase-3 and caspase-9, and suppressed nuclear factor (NF)-kappaB expression in tumor tissue as determined by reverse transcription (RT)-PCR and western blotting. In summary, shRNA targeting of E2F-1 can effectively inhibits human gastric cancer MGC-803 cell growth in vivo and may be a potential therapeutic strategy for gastric cancer.
Animals ; Apoptosis ; Blotting, Western ; Caspase 3/metabolism ; Caspase 9/metabolism ; Cells, Cultured ; E2F1 Transcription Factor/antagonists & inhibitors/genetics/*metabolism ; Genetic Vectors/administration & dosage ; Humans ; Lentivirus/*genetics ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; RNA Interference ; RNA, Small Interfering/*genetics ; Stomach Neoplasms/*genetics/pathology/*prevention & control

The death of patients with gastric cancer is mainly due to its recurrence and metastasis, and circulating tumor cell (CTC) is the necessary condition of metastasis. As liquid biopsy, CTC detection has its certain clinical significance. The detection is required after enrichment because circulating tumor cells are rare. Many enrichment methods have been developed: methods based on physical characteristics of TCT, like density, size and dielectric properties and so on; immunogenicity, like Cell Search System; and microfluidic chip technology. The immunofluorescence is commonly used to identify CTC in gastric cancer and the isolated CTC can also be used for the following analysis on the level of nucleic acid, protein and gene regulation. Detection of CTC in gastric cancer is helpful to judge the prognosis, assess staging, monitor the curative effect and guide the development of drug. There are many challenges for clinical transformation of CTC: the lower enrichment efficiency, the less specific surface markers, the uncertain diagnostic efficiency and so on, but it also has the good research prospect because it is non-invasive, repeatable and can real-time monitor the condition and guide the clinical treatment compared with pathological biopsy. In this paper, the detection and identification methods, and clinical value of CTC in gastric cancer patients are reviewed.
Biomarkers, Tumor ; Biopsy ; Cell Separation ; methods ; Cytodiagnosis ; methods ; Flow Cytometry ; methods ; Fluorescent Antibody Technique ; methods ; Humans ; Microchip Analytical Procedures ; methods ; Neoplasm Recurrence, Local ; prevention & control ; Neoplasm Staging ; methods ; Neoplastic Cells, Circulating ; metabolism ; pathology ; Prognosis ; Secondary Prevention ; Stomach Neoplasms ; blood ; diagnosis ; genetics ; therapy ; Treatment Outcome