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

OBJECTIVETo investigate the expression of Versican in gastric carcinoma and its relationship with tumor angiogenesis.

METHODSProtein expression of Versican, vascular endothelial growth factor and CD34 was evaluated by immunohistochemistry (EliVision method) in 80 cases of gastric carcinoma and 30 samples of normal gastric tissue.

RESULTSThere were statistically significant differences in the expression of Versican, vascular endothelial growth factor and CD34 between gastric carcinoma and normal gastric tissue (P < 0.05). The expression of Versican was seen mainly in fibroblasts of the tumor and was correlated with tumor differentiation, clinical stage and lymph node metastasis (P < 0.05), whereas vascular endothelial growth factor was primarily seen in the cytoplasm of the tumor cells and correlated with tumor differentiation, clinical stage, Lauren classification and lymph node metastasis (P < 0.05). MVD was correlated with tumor differentiation, clinical stage, Lauren classification, depth of tumor invasion and lymph node metastasis (P < 0.05). In addition, positive correlation of Versican and VEGF protein expression was found in tumor cells (r = 0.467, P < 0.01).

CONCLUSIONThe expression of both Versican and vascular endothelial growth factor is closely associated with tumor angiogenesis in gastric carcinoma.

Antigens, CD34 ; metabolism ; Fibroblasts ; metabolism ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Neoplasm Proteins ; metabolism ; Neovascularization, Pathologic ; Prognosis ; Stomach ; metabolism ; Stomach Neoplasms ; blood supply ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; metabolism ; Versicans ; metabolism

OBJECTIVETo explore the effect and mechanism of autophagy specific gene Beclin-1 in gastric cancer cell SGC7901 on vasculogenic mimicry (VM) forming ability.

METHODSPlasmid vectors with and without integrated shRNA were transfected respectively into SGC7901 cell line (Beclin1-inhibited group and negative control group). Simple SGC7901 cell line was used as blank group. RT-PCR and Western blot were performed to examine the expression of Beclin-1 in 3 groups. Culture was used to construct the VM model in vitro. Different VM forming ability was measured and genes (beclin-1, notch-1) expression of each group was detected before and after VM formation.

RESULTSBeclin-1 and notch-1 expression increased significantly in the process of VM forming. When beclin-1 was inhibited, the formation of VM was limited and VM formative genes expression decreased. As compared to cells of negative control group, those of Beclin1-inhibited group had less number of VM forming cellular tube-like construction (15.4±1.1 vs. 37.8±1.9, P<0.05), shorter length of such construction [(316.8±24.6) mm vs. (385.1±14.2) mm, P<0.05], and less crossing point (11.6±1.1 vs. 27.2±1.1, P<0.05).

CONCLUSIONSBeclin-1 can promote VM formation through maintaining stable expression of gastric cancer cell VM regulating genes. Beclin-1 inhibition may be a new target for gastric cancer gene therapy.

Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; Beclin-1 ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Membrane Proteins ; metabolism ; RNA, Small Interfering ; Stomach Neoplasms ; blood supply ; metabolism ; Transfection

OBJECTIVETo explore the possible angiogenesis mechanism of Huazhuo Jiedu Hewei Recipe (HJHR) in preventing and treating precancerous lesions of gastric cancer (PLGC).

METHODSTotally 66 Wistar rats were randomly divided into 6 groups, i.e., the normal control group, the model group, the retinoic acid (RA) group, the high dose HJHR group, the middle dose HJHR group, the low dose HJHR group, 11 in each group. PLGC model was duplicated by inserting a spring with Helicobacter. Corresponding medicines were administered to rats in each medicated group once daily by gastrogavage, 2 mL each time for 12 successive weeks. The effect of HJHR on hypoxia induced factor (HIF-1alpha) and vascular endothelial growth factor (VEGF) of PLGC in chronic atrophic gastritis (CAG) rats' gastric mucosa was observed by immunohistochemical assay and Western blot method.

RESULTSCompared with the normal control group, the expression of VEGF and HIF-1alpha increased in the model group (P < 0.05). Compared with the model group, the expression of VEGF and HIF-1alpha decreased in each medicated group (P < 0.05). Besides, they were lower in the high and middle dose HJHR groups than in the RA group and the low dose HJHR group (P < 0. 05). There was no statistical difference between the low dose HJHR group and the RA group (P > 0.05).

CONCLUSIONHJHR could prevent and treat PLGC of CAG rats possibly through decreasing the expression of HIF-1alpha and VEGF in a dose-dependent manner.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; metabolism ; Gastritis ; metabolism ; microbiology ; Helicobacter ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Neovascularization, Pathologic ; Precancerous Conditions ; blood supply ; drug therapy ; metabolism ; Rats ; Rats, Wistar ; Stomach Neoplasms ; blood supply ; drug therapy ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

The development of gastric cancer (GC) is closely related to chronic inflammation caused by Helicobacter pylori infection, and herpes virus entry mediator (HVEM) is a receptor expressed on the surface of leukocytes that mediates potent inflammatory responses in animal models. However, the role of HVEM in human GC has not been studied. Previously, we showed that the interaction of HVEM on human leukocytes with its ligand LIGHT induces intracellular calcium mobilization, which results in inflammatory responses including induction of proinflammatory cytokine production and anti-bacterial activities. In this study, we report that leukocytes from GC patients express lower levels of membrane HVEM (mHVEM) and have lower LIGHT-induced bactericidal activities than those from healthy controls (HC). In contrast, levels of soluble HVEM (sHVEM) in the sera of GC patients were significantly higher than in those of HC. We found that monocyte membrane-bound HVEM is released into the medium when cells are activated by proinflammatory cytokines such as TNF-alpha and IL-8, which are elevated in the sera of GC patients. mHVEM level dropped in parallel with the release of sHVEM, and release was completely blocked by the metalloprotease inhibitor, GM6001. We also found that the low level of mHVEM on GC patient leukocytes was correlated with low LIGHT-induced bactericidal activities against H. pylori and S. aureus and production of reactive oxygen species. Our results indicate that mHVEM on leukocytes and sHVEM in sera may contribute to the development and/or progression of GC.
Aged ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Monocytes/metabolism ; Neutrophils/metabolism ; Reactive Oxygen Species/metabolism ; Receptors, Tumor Necrosis Factor, Member 14/*blood ; Stomach Neoplasms/*blood/metabolism ; Tumor Necrosis Factor Ligand Superfamily Member 14/blood

OBJECTIVETo study the inhibition and killing effect of transgenic LIGHT umbilical cord blood mesenchymal stem cells (UCBMSCs) on stomach carcinoma.

METHODSThe LIGHT gene was recombined to construct the transfer plasmid pGC-FU-LIGHT by infusion technique. The 293T cells were co-transfected with the transfer plasmid pGC-FU-LIGHT, the construction plasmid Helper 1.0 and the envelope plasmid Helper 2.0 with the help of lipofectamine 2000 to produce lentiviral particles. Transgenic UCBMSCs(MSC-LIGHT) and empty carrier UCBMSCs (MSC) were obtained. Human gastric cancer cell SGC-7901 was injected into nude mice subcutaneously groin. The model of transplanted human gastric cancer cell SGC-7901 in nude mice was established. Tumorigenesis nude mice were separated into three groups randomly with 5 in each group: MSC-LIGHT group, MSC group, and NS group. Three groups of nude mice were injected around the tumor with MSC-LIGHT, MSC and NS every other day for 3 times. Four weeks later, the transplanted gastric cancer volume was measured. The expressions of LIGHT in the three groups were determined by RT-PCR and ELISA method. The necrosis area in the tumors was calculated under pathological examination.

RESULTSThe average volume of transplanted tumor was(0.45±0.25) cm(3) in MSG-LIGHT group, (0.64±0.36) cm(3) in MSG group, and(1.21±0.79) cm(3) in NS group, and the difference was statistically significant(P<0.05). The LIGHT mRNA was 2.96±0.27, 1.23±0.47, and 0.73±0.10 respectively. The LIGHT protein was(167.89±2.31), (73.22±5.74), and (49.66±5.25) ng/L. The differences were all statistically significant among the three groups(both P<0.01). Pathological examination showed that the necrosis area was largest in MSC-LIGHT group.

CONCLUSIONTransgenic UCBMSCs secret LIGHT in a paracrine manner, which has inhibition and killing effects on stomach carcinoma.

Animals ; Cell Line, Tumor ; Fetal Blood ; cytology ; Genetic Therapy ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Plasmids ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; therapy ; Transfection ; Tumor Necrosis Factor Ligand Superfamily Member 14 ; genetics ; metabolism ; Xenograft Model Antitumor Assays

The study is to investigate the pharmacokinetics of S-1 capsule (tegafur, gimeracil and potassium oxonate capsule) in patients with advanced gastric cancer after single and multiple oral administration. Twelve patients with advanced gastric cancer were recruited to the study. The dose of S-1 for each patient was determined according to his/her body surface area (BSA). The dose for single administration was 60 mg every subject. The dose for multiple administration for one subject was as follows: 100 mg x d(-1) or 120 mg x d(-1), 28-days consecutive oral administration. The pharmacokinetic parameters of tegafur, 5-fluorouracil, gimeracil, potassium oxonate and uracil after single oral administration were as follows: (2,207 +/- 545), (220.0 +/- 68.2), (374.9 +/- 103.0), (110.5 +/- 100.8) and (831.1 +/- 199.9) ng x mL(-1) for Cmax; (11.8 +/- 3.8), (4.4 +/- 3.3), (7.8 +/- 5.1), (3.1 +/- 0.9) and (8.8 +/- 4.1) h for t1/2, respectively. After six days oral administration, the average steady state plasma concentrations (Cav) of tegafur, 5-fluorouracil, gimeracil, potassium oxonate and uracil were (2,425 +/- 1,172), (73.88 +/- 18.88), (162.6 +/- 70.8), (36.89 +/- 29.35) and (435.3 +/- 141.0) ng x mL(-1), respectively, and the degree of fluctuation (DF) were (1.0 +/- 0.2), (2.5 +/- 0.4), (3.1 +/- 0.8), (2.4 +/- 0.8) and (1.5 +/- 0.3), respectively. The cumulative urine excretion percentage of tegafur, 5-fluorouracil, gimeracil and potassium oxonate in urine within 48 h were (4.2 +/- 2.8) %, (4.7 +/- 1.6) %, (18.5 +/- 6.0) % and (1.7 +/- 1.2) %, repectively, after single oral administration of S-1. The results exhibited that tegafur had some drug accumulation observed, and gimeracil, potassium oxonate, 5-fluorouracil and uracil had no drug accumulation observed.
Administration, Oral ; Adult ; Aged ; Antimetabolites, Antineoplastic ; pharmacokinetics ; Capsules ; Drug Combinations ; Female ; Fluorouracil ; blood ; urine ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Oxonic Acid ; blood ; pharmacokinetics ; urine ; Pyridines ; blood ; urine ; Stomach Neoplasms ; blood ; metabolism ; pathology ; urine ; Tegafur ; blood ; pharmacokinetics ; urine ; Uracil ; blood ; urine

OBJECTIVETo explore the role of oxidative stress and the antioxidant protein thioredoxin in the tumorigenesis and progression of gastric cancer.

METHODSThe plasma levels of adenosine deaminase (ADA), glutathione peroxidase (GPX), superoxide dismutase (SOD), and advanced oxidation protein products (AOPP) were determined by colorimetry, and the plasma levels of thioredoxin were determined by enzyme-linked immunosorbent assay (ELISA) in 48 gastric cancer patients and 30 healthy subjects. RT-PCR assay was employed to examine the expression levels of thioredoxin mRNA in the tissue samples of the patients.

RESULTSCompared with the healthy controls, patients with gastric cancer had significantly increased plasma levels of ADA and AOPP (P<0.05), decreased plasma GPX level (P<0.05), and similar plasma SOD levels. The plasma levels of thioredoxin were significantly higher in patients with gastric cancer than in the healthy controls (P<0.05). Thioredoxin levels was not associated with gender, age, degree of tumor cell differentiation, invasion depth, or lymph node metastasis (P>0.05), but was correlated to distant tumor metastasis (P<0.05). The expression of Trx mRNA was significantly higher in gastric carcinoma than in normal gastric tissue (P<0.05).

CONCLUSIONGastric cancer patients have high levels of oxidative stress and thioredoxin expression, and the latter is related to distant metastasis of the tumor.

Adenosine Deaminase ; blood ; Adult ; Advanced Oxidation Protein Products ; blood ; Aged ; Case-Control Studies ; Female ; Glutathione Peroxidase ; blood ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Oxidative Stress ; RNA, Messenger ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Superoxide Dismutase ; blood ; Thioredoxins ; genetics ; metabolism

OBJECTIVETo evaluate the prognostic significance of preoperative serum albumin in patients with gastric cancer undergoing radical resection.

METHODSA total of 146 patients with gastric cancer underwent radical resection from January 2001 to December 2003. Clinicopathological data were analyzed retrospectively. Patients were divided into two groups, including patients with a normal preoperative serum albumin level(>35 g/L, n=115) and patients with hypoalbuminemia (≤35 g/L, n=31).

RESULTSPatients with a low albumin level were associated with a higher postoperative recurrence rate(90.3% vs. 43.5%, P<0.01). The overall 5-year survival rate in patients with a normal serum albumin level was significantly higher than that in patients with a low serum level(57.4% vs. 9.7%, P<0.01). On multivariate analysis, preoperative serum albumin level was an independent factor associated with survival(P<0.01). When stratified by nodal metastasis, normal serum albumin level was still associated with higher survival rate(P<0.05). Prognostic significance was found in patients with lower stomach cancer(P<0.01), but not in patients with cancer in the upper and middle stomach(P>0.05).

CONCLUSIONHypoalbuminemia is associated with worse survival in patients with cancer in the lower stomach and adjuvant therapy should be considered.

Adult ; Aged ; Chi-Square Distribution ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Serum Albumin ; metabolism ; Stomach Neoplasms ; blood ; surgery

OBJECTIVETo establish experimental models for tumor neovascularization and to apply quantitative digital imaging analysis in the study.

METHODSAn endothelial tube formation model was established by human umbilical vein endothelial cells (HUVECs). A vasculogenic mimicry model was established by SGC-7901 gastric cancer cell line. Fertilized eggs were used to establish a chorioallantoic membrane angiogenesis model. Using gene transfection experiment, IRX1 tumor suppressor gene was chosen as a therapeutic target. Image Pro Plus (IPP) analysis software was used for digital vascular images analysis with parameters including points, lines, angles and integral absorbance (IA) for the tubular formation or vasculogenic mimicry.

RESULTSDigital image analysis by IPP showed that HUVEC tubular formation was significantly inhibited in IRX1 transfectant, compared with controls. The tubular numbers in three groups were 12.80 +/- 3.83, 29.00 +/- 5.34 and 28.20 +/- 4.32 (P<0.01). The connection points of tubules in three groups were 13.20 +/- 2.59, 25.00 +/- 2.24 and 24.60 +/- 3.21 (P<0.01). The tubular lengths of three groups were (821.5 +/- 12.5), (930.9 +/- 13.5) and (948.4 +/- 18.1) microm (P=0.022). The IA values of PAS stain in three groups were 3606 +/- 363, 14 200 +/- 1251 and 15 043 +/- 1220 (P<0.01). In chick chorioallantoic membrane model, the angular numbers of tubules in three groups were 6.41 +/- 2.60, 10.27 +/- 2.65 and 9.18 +/- 1.99 (P<0.01).

CONCLUSIONSThe endothelial tube formation model, vasculogenic mimicry model and chorioallantoic membrane angiogenesis model are useful for gene therapy and drug screening with targeting neoplastic vascularization. Professional image analysis software may greatly facilitate the quantitative analysis of tumor neovascularization.

Animals ; Cell Line, Tumor ; Cells, Cultured ; Chorioallantoic Membrane ; blood supply ; Diagnostic Imaging ; methods ; Homeodomain Proteins ; genetics ; metabolism ; physiology ; Human Umbilical Vein Endothelial Cells ; Humans ; Neovascularization, Pathologic ; Neovascularization, Physiologic ; Software ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transcription Factors ; genetics ; metabolism ; physiology ; Transfection