Death In Gastric Cancer Patients Research Articles

Milky spot macrophages remodeled by gastric cancer cells promote peritoneal mesothelial cell injury

Peritoneal dissemination (PD) is the most frequent metastatic pattern of advanced gastric cancer (GC) and the main cause of death in GC patients. Human peritoneal mesothelial cell (HPMC) injury induced by gastric cancer cells (GCCs) and GCC outgrowths supported by peritoneal milky spot macrophages (PMSMs) are the key events during gastric cancer peritoneal dissemination (GCPD). In this study, we investigated whether PMSMs remodeled by GCC can induce HPMC injury and create a favorable microenvironment for GCPD. We established a tumor-associated macrophage (TAM) model using in vitro cell coculture. Normal macrophages cocultured with GCCs down-regulated expression of antigen-presenting surface molecules CD80, CD86, and MHC-II, but, notably, they up-regulated expression of phagocytic scavenger receptor CD206, which is similar to the M2 macrophage phenotype. In further experiments, various experimental methods were applied to detect the injurious effect of TAMs on HPMCs in another TAM–HPMC coculture. Our results showed that GCCs can induce HPMC apoptosis by unregulated apoptosis associated with cleaved caspase3, cleaved caspase9, and p21 proteins. HPMC growth ceased, and both early- and late-stage apoptosis were observed. Additionally, GCCs can induce HPMC fibrosis via increased expression of epithelial cell marker E-cadherin and decreased expression of mesenchymal cell marker α-SMA. Our results demonstrate that, in the GCPD process, PMSMs were remodeled by GCCs, resulting in phenotypic and functional transformation. In turn, this transformation induced HPMC injury and provided a favorable microenvironment for GCC anchorage and growth. These results may provide new insight into the mechanisms of GCPD.

The cytotoxic effect of TGF-β1 on mesothelial cells via apoptosis in early peritoneal carcinomatosis

Peritoneal dissemination is one of the main causes of death in gastric cancer patients. We have previously reported that gastric cancer cells can induce peritoneal apoptosis, lead to damage of peritoneum integrity, and therefore promote peritoneal metastasis. However, the soluble factors secreted by cancer cells to trigger the damaging cascade remain unclear. TGF-β1, a cytokine known for its capacity to induce proliferative and transformative changes of cells is found in significantly higher quantities correlated with peritoneal metastasis and TNM stages of gastric cancer. High levels of TGF-β1 in the subperitoneal milieu may affect the morphology and function of mesothelial cells, so that the resulting environment becomes favorable for peritoneal metastases. We observed apoptosis induced by TGF-β1 in mesothelial cells in peritoneal carcinomatosis. Knockdown of the smad2 gene by siRNA silencing can partially inhibit these effects. TGF-β1 could upregulate the expressions of Bax and suppress Bcl-2 in mesothelial cells. We conclude that TGF-β1 could induce apoptosis of mesothelial cells, which involves the smad2 signaling pathway in peritoneal carcinomatosis. Bcl-2 and Bax may contribute to this phenomenon.

TGF-β1 induces peritoneal fibrosis by activating the Smad2 pathway in mesothelial cells and promotes peritoneal carcinomatosis.

Peritoneal dissemination is one of the main causes of death in gastric cancer patients. Our previous study demonstrated that peritoneal fibrosis induced by transforming growth factor-β1 (TGF-β1) may provide a favorable environment for the dissemination of gastric cancer. The role of Smad3 in the development of dermal fibrosis, subcapsular cataract, and peritoneal fibrosis has been reported. However, the potential role of Smad2 in the development of fibrosis is unclear. The objective of this study was to determine the effect of Smad2 in peritoneal fibrosis, induced by TGF-β1, on dissemination of gastric cancer. Here we demonstrate that TGF-β1 significantly stimulated the expression of collagenIII and fibronectin in mesothelial cells through the Smad2 signal transduction pathway, but knockdown of the Smad2 gene by silencing siRNA partially inhibited these effects. This inhibition was associated with a depressed adhesion and invasiveness of gastric cancer cells. We conclude that peritoneal fibrosis induced by TGF-β1 is dependent on Smad2 signaling and may provide a hospitable environment for carcinomatosis.

Expression and clinical significance of CD8 + T cells in gastric carcinoma tissues

Objective To study the expression and clinical significance of infiltrating lymphocytes CD8 + T in gastric carcinoma tissues.Methods The expression of infiltrating lymphocytes CD8 + T in 102surgically resected specimens of gastric cancer tissues was detected by using immunohistochemistry.Results CD8+ T cells were expressed both in gastric carcinoma,gastritis and gastric polyp tissues,but the number of CD8 + T cells in gastric carcinoma tissues [(29.53 ± 14.71 )/high powered field (HPF)]wassignificantly greater than in gastritis tissues [( 19.54 ±8.10)/HPF]and polyp tissues [( 14.26 ±6.83)/HPF](P ＜0.05).No correlation was found between CD8 + T lymphocyte count and gender,age,tumor location,tumor size,lymph node metastasis,tumor relapse,pathological grade or TNM stage ( P ＞ 0.05 ).However,there was a statistically significant correlation between the number of CD8 + T cells and the histological type,invasive depth.The CD8 + T cell count was obviously higher in the differentiated group [(33.9 ± 18.4)/HPF]than in the undifferentiated group [(27.6 ± 12.0)/HPF]( t = 2.04,P＜0.05 ).And the number of CD8 + T cells was greater in the non-invasive group [(38.8 ± 13.8)/HPF]than in the invasive group [(27.7 ± 14.1 )/HPF]( t = 2.45,P＜0.05 ).The median survival time of gastric cancer patients with high expression of CD8 + T cells was 5 months longer than in those with low expression,but there was no significant difference.Multi-factor COX model analysis demonstrated that the risk of death in gastric cancer patients was lower in the CD8 + T cells high expression group than in the low expression group ( RR = 0.82,95%,CI = 0.41-1.65 ).Conclusion Detection of infiltrating lymphocytes CD8 + T expression in gastric carcinoma tissues will be beneficial to the judgment of the prognosis. Key words: Gastric carconoma; Infiltrate; Lymphocytes; CD8

Human peritoneal mesothelial cell transformation into myofibroblasts in response to TGF-ß1 in vitro

Peritoneal dissemination is one of the leading causes of death in gastric cancer patients. The interaction between carcinoma cells and the peritoneal lining may play a key role in tumor peritoneal dissemination. Human peritoneal mesothelial cells are a monolayer of squamous epithelial cells covering the peritoneal cavity and forming serosal membranes. The precise role of mesothelial cells in the peritoneal dissemination of gastric cancer remains to be identified. Expression of TGF-ß1, a cytokine known for its capacity to induce proliferative and transformative changes in cells, has been correlated with peritoneal metastasis and TNM stages of gastric cancer. High levels of TGF-ß1 in the subperitoneal milieu may play a key role in the transition of normal mesothelial cells to myofibroblasts. Here, we demonstrate that mesothelial cells activated by TGF-ß1 undergo epithelial-mesenchymal transition (EMT) and that the transition of mesothelial cells to myofibroblasts is dependent on Smad2 signaling. EMT of mesothelial cells was marked by up-regulation of α-smooth muscle actin and vimentin expression. Cytokeratin and E-cadherin expression decreased over time in transformed mesothelial cells. Knockdown of Smad2 gene by siRNA silencing significantly suppressed the transition of mesothelial cells to myofibroblasts. We conclude that when exposed to TGF-ß1 mesothelial cells undergo EMT which involves Smad2 signaling. Furthermore, mesothelial cells may be the possible source of myofibroblasts in peritoneal fibrosis and provide a favorable environment for the dissemination of gastric cancer.

Induction of gastric cancer cell adhesion through transforming growth factor-beta1-mediated peritoneal fibrosis

BackgroundPeritoneal dissemination is one of the main causes of death in gastric cancer patients. Transforming growth factor-beta1 (TGF-β1), one of the most potent fibrotic stimuli for mesothelial cells, may play a key role in this processing. The purpose of this study is to elucidate the effects of TGF-β1 on regulation of gastric cancer adhesion to mesothelial cells.MethodsPeritoneal tissues and peritoneal wash fluid were obtained for hematoxylin and eosin staining or ELISA to measure fibrosis and TGF-β1 levels, respectively. The peritoneal mesothelial cell line, HMrSV5, was used to determine the role of TGF-β1 in regulation of gastric cancer cell adhesion to mesothelial cells and expression of collagen, fibronectin, and Smad 2/3 by using adhesion assay, western blot, and RT-PCR.ResultsThe data showed that TGF-β1 treatment was able to induce collagen III and fibronectin expression in the mesothelial cells, which was associated with an increased adhesion ability of gastric cancer cells, but knockdown of minimal sites of cell binding domain of extracellular matrix can partially inhibit these effects.ConclusionPeritoneal fibrosis induced by TGF-β1 may provide a favorable environment for the dissemination of gastric cancer.

Multi-factor COX model analysis of impact of costimulatory molecules B7-H4 on gastric cancer prognosis

Objective To study the relation of B7-H4 expression and gastric cancer, and to analyze the impact of different expression levels of costimulatory molecules B7-H4 in gastric cancer tissues on survival time of gastric cancer patients treated with cytokine-induced killer cell biotherapy. Methods 156 gastric cancer patients who received operative treatment and were diagnosed pathologically in the Third Affiliated Hospital, Soochow University, were followed up from Jan. 1, 1998 to Dec. 31,2009. The B7-H4 expression was detected by using immunohistochemical staining and IHS assessment analysis. Demngraphic data, clinical data, recurrence and death time of gastric cancer patients were collected by the method of retrospective cohort study. Impact of B7-H4 high expression on the risk of gastric cancer recurrence and death was analyzed through multi-factor COX model. Results In B7-H4 high expression group and B7-H4 low expression group, the median tumor-free survival time (months) of gastric cancer patients and 95% CI of gastric cancer patients was respectively 16 ( 11.9-20. 1 ) and 47 ( 22.4-71.6 ), and 25 ( 19.0-31.1 )and 43 (35. 2-50. 8). In the chemotherapy group, the median survival time of gastric cancer patients in B7-H4 low expression group was significantly longer than that in B7-H4 high expression group ( 36 vs 16 ,x2 = 14. 14,P 0. 05). In the chemotherapy group, the median tumor-free survival time of gastric cancer patients in B7-H4 low expression group was significantly longer than that in B7-H4 high expression group (33 vs 11, x2= 18. 956,P <0. 01 ). In the chemotherapy plus CIK treatment group, the median tumor-free survival time of gastric cancer patients in B7-H4 low expression group was significantly longer than that in B7-H4 high expression group (90 vs 18 ,x2= 3. 842, P < 0. 05 ). After confounding factors such as sex, age and so on were adjusted, risk of recurrence and death of gastric cancer patients in B7-H4 high expression group was significantly higher than in B7-H4 low expression group ( RR =2. 30, 95% CI = 1.41-3. 75; RR = 1.90, 95% CI =1.20-3.01, respectively). Conclusion B7-H4 high expression may be an independent risk factor of increasing the risk of gastric cancer recurrence and death. CIK cell transfusion treatment can control and interfere in the B7-H4 expression, and prolong the median survival time of gastric cancer patients. B7-H4 low expression of gastric cancer can be considered as inclusion criteria of CIK cell biotherapy. Key words: Stomach carcinoma; B7-H4; Costimulatory molecules; Immunotherapy; Survival time

Increasing the frequency of CIK cells adoptive immunotherapy may decrease risk of death in gastric cancer patients

to analyze the correlation between cytokine-induced killer (cik) cells adoptive immunotherapy and cancer-related death in gastric cancer patients. methods: One hundred and fifty-six gastric cancer patients after operation at the Third Affiliated Hospital of Soochow University were enrolled in this study. Their clinical data including demographic characteristics, operation time, tumor size, pathological type and staging, tumor metastasis, outcome of chemotherapy or CIK cells adoptive immunotherapy, survival time or time of death were collected with a standard structured questionnaire. Kaplan-Meier method was used to estimate the median survival time, and the 2- and 5- year survival rates. Hazard risk (HR) and 95% confidence interval (95% CI) of CIK cells adoptive immunotherapy for gastric cancer were calculated using the two-stage time-dependent covariates Cox model. the survival time of gastric cancer patients was longer after CIK cells adoptive immunotherapy than after chemotherapy (χ(2) = 10.907, P = 0.001). The median survival time of gastric cancer patients was also longer after CIK cells adoptive immunotherapy than after chemotherapy (49 mo vs 27 mo, P < 0.05). The 2- and 5-year survival rates of gastric cancer patients were significantly higher after CIK cells adoptive immunotherapy than after chemotherapy (73.5% vs 52.6%, 40.4% vs 23.9%, P < 0.05). A significant difference was observed in the survival curve for patients who received CIK cells adoptive immunotherapy (0, 1-10, 11-25, and over 25 frequencies) (χ(2) = 14.534, P = 0.002). The frequencies of CIK cells adoptive immunotherapy were significantly related with the decreasing risk of death in gastric cancer patients after adjustment for sex and age of the patients, tumor stage and relapse (HR = 0.54, 95% CI: 0.36-0.80) when the first stage Cox model was used to define the subjects who remained alive beyond 36 mo as survivors. However, no correlation was observed between the frequencies of death in CIK cells adoptive immunotherapy and the risk of gastric cancer patients (HR = 1.09, 95% CI: 0.63-0.89) when the second stage Cox model was used to define the subjects who survived for more than 36 mo as survivors. the survival time of the gastric cancer patients treated with chemotherapy combined with CIK cells adoptive immunotherapy is significantly longer than that of the patients treated with chemotherapy alone and increasing the frequency of CIK cells adoptive immunotherapy seems to benefit patients more.

Expression of costimulatory molecule B7-H4 in the gastric carcinoma tissues and its correlation with prognosis

Objective To study the expression of eostimulatory molecules BT-H4 in the gastric carcinoma tissues and its correlation with prognosis.Methods B7-H4 expression in the surgical specimens of gastric cancer tissues was detected by using immunohistochemistry in 156 patients who had obtained either autologous cytokine-induced killer(CIK)cell biotherapy or chemotherapy.Results Positive expression rate of B7-H4 in gastric carcinoma tissues was 44.9%.Single-factor analysis demonstrated that there was no correlation between the B7-H4 expression and sex,age,histological type,pathological grade or tumor size.However,there was a statistically significant correlation between the BT-H4 positive expression and the invasion depth and lymph node metastasis.The positive expression rate of B7-H4 was obviously higher in the invasive group than in the non-invasive group(49.1% vs 16.7%,X2 = 8.467,P ＜ 0.01).And the positive expression rate of B7-H4 was higher in the lymph node metastasis group than in the nonmetastasis group(X2 = 17.339,P ＜0.01).The median survival time of gastric cancer patients with B7-H4 low expression was 13 months longer than in those with high expression(X2 = 12.38,P ＜0.01).Multi-factor COX model analysis demonstrated that the B7-H4 expression may be considered as the prediction of survival time in the gastric cancer patients(RR=1.85,95% CI = 1.15-2.96).Moreover,the risk of death in gastric cancer patients was significantly lower in the CIK treatment group than in the chemotherapy group(RR =0.53,95% CI =0.33-0.85).Conclusion There was a negative correlation between the high expression of B7-H4 and the survival time of gastric cancer patients,which may suggest that B7-H4 is one of negative regulatory molecules and may be considered as a predictive index of gastric cancer patients.CIK cells treatment can significantly prolong the survival time. Key words: Stomach carcinoma; B7-H4; Immunotherapy; Immunohistochemical; Costimulatory molecule

Lymph node ratio and survival for gastric cancer: A retrospective analysis of the SEER database

4561 Purpose: To determine the prognostic significance of the lymph node ratio (ratio of number of positive lymph nodes to number of dissected lymph nodes) in gastric cancer patients. Methods: We retrospectively analyzed 10,176 gastric patients from 1990–2003 who underwent curative gastrectomy from the SEER database. Survival curves were calculated according to the Kaplan-Meier method and analyzed with log-rank test. Multivariate analysis of prognostic factors related to survival was performed by the Cox proportional hazard model. Results: The lymph node ratio (LNR) was a strong predictor of survival. LNR was equally predictive of survival whether the analysis was restricted to patients with &lt;15 lymph nodes dissected or &gt;15 lymph nodes dissected. Survival of patients with a LNR of 0.1–5% was not significantly different than node negative patients; however, survival of patients with a LNR of 5–10% was significantly different than node negative patients. Multivariate analysis showed that LNR, T-stage, tumor size, and number of lymph nodes positive were independent prognostic predictors of death and that LNR was the strongest predictor for death. Multivariate analysis showed that the number of lymph nodes dissected was an independent prognostic factor for survival. Moreover, LNR was an independent prognostic factor for N1 and N2 patients by AJCC staging. LNR trended toward significance in AJCC N3 patients. Conclusions: LNR was the strongest predictor of death in gastric cancer patients when compared to T-stage, number of lymph nodes positive, and tumor size. LNR is equally predictive regardless of the adequacy of the lymph node dissection. No significant financial relationships to disclose.

Gastric cancer cell detection in peritoneal lavage: RT-PCR for carcinoembryonic antigen transcripts versus the combined cytology with peritoneal carcinoembryonic antigen levels

Free cancer cells exfoliated from the serosal surfaces of primary cancers are considered to be responsible for peritoneal dissemination, which are both the most frequent pattern of treatment failure and the most important cause of death in gastric cancer patients. Detection of free cancer cells in the peritoneal cavity at the time of surgery, therefore, is thought to be of great value in predicting peritoneal recurrence and accordingly the prognosis of gastric cancer patients. This study was designed to determine whether free cancer cells in peritoneal lavage fluid from gastric cancer patients could be identified by a reverse-transcriptase polymerase chain reaction (RT-PCR) method specific to carcinoembryonic antigen (CEA) mRNA. Simultaneously, the results from conventional cytological examination were evaluated and the levels of CEA in peritoneal lavage were determined. Of the 40 gastric cancer patients enrolling in this investigation, 11 (27.5%) were positive for CEA mRNA in their peritoneal lavage, whereas only 6 (15%) and 8 (20%) were shown to be positive by cytological examination and peritoneal CEA (pCEA) assay, respectively. Furthermore, RT-PCR positive for CEA mRNA was correlated with the depth of tumor invasion ( P<0.001), lymph node metastases ( P=0.004), the TNM stage ( P<0.001) and peritoneal recurrence ( P<0.001). The technique of RT-PCR was more sensitive than conventional cytological examination and pCEA levels in the detection of peritoneal free cancer cells as well as in the prediction of peritoneal recurrence. In addition, CEA RT-PCR had a high concordance rate (82.5%) with the combination of cytology with pCEA levels. These observations suggest that it is feasible to identify free cancer cells in peritoneal lavage by using a CEA mRNA-specific RT-PCR method, and this assay can be a promising diagnostic modality for evaluating the risk of peritoneal dissemination in gastric cancer patients following operations.