Objective To find out some ideal reconstructions after total gastrectomy in experimental study of rat. Methods Sixty male Sprague-Dawley rats were randomly and averagely divided into 6 groups: Roux-en-Y group (RY group), proximate jejunal pouch group (PJP group), distal jejunal pouch group (DJP group), two jejunal pouchs group (TJP group), duodenumjejunal pouch interposition group (DJPI group) and laparotomy group (L group). Body weight of rats, intestinal transit distance, adaptive changes in esophagojejunostomic mucosa and morphology changes of intestine after operation were observed and compared. Results At 2 weeks after operation, body weight in each group were significantly lower than that before operation （P＜0.05）. At 4 weeks postoperatively, body weight in PJP group, TJP group and DJPI group were significantly higher than that in RY group respectively （P＜0.05）, as well as at 8 weeks. Intestinal transit distance in PJP group was shorter than that in RY group （P＜0.05）. With regard to intestinal mucosa, TJP group and DJPI group were significantly different with RY group （P＜0.05）. Interestingly, there was no difference in each group as to refluxing esophagitis （P＞0.05）. Conclusion Proximate and two jejunal pouchs Roux-en-Y esophagojejunostomy seem to be ideal procedures for digestive tract reconstruction after total gastrectomy. The jejunal pouch interposition procedure seems to be same effective to PJP and TJP, but there is no preponderance over the former.
ObjectiveTo detect the expressions of CD133 in four gastric cancer cell lines (KATO-Ⅲ, SGC7901, AGS, and MKN-45) and investigate the different biological characteristics of CD133 positive (CD133+) cells and CD133 negative (CD133-) cells in the KATO-Ⅲ cell lines. MethodsThe CD133 gene and protein expressions of four gastric cancer cell lines were evaluated by semiquantitative RT-PCR and Western blot, respectively. CD133+ cells in KATO-Ⅲ cell lines were isolated by magnetic activated cell sorting (MACS) and examined in morphology, growth characteristics, proliferation, and differentiation in vitro. The sensitivity of different concentrations (0.05, 0.10, 0.20, 0.50, and 1.00 mg/ml) 5-fluoropyrimidinedione (5-FU) were contrasted by drug sensitivity testing in vitro (Cell Counting Kit 8-assay) between CD133+ cells and CD133- cells. ResultsThe expressions of CD133 gene and protein in the KATO-Ⅲ cell lines were significantly higher than those in the other three cell lines (Plt;0.05). CD133+ cells produced spheroid colonies in serum-free medium culture and were ber abilities of proliferation and differentiation than those of CD133- cells in vitro. The inhibitor rate of the CD133+ cells at concentration of 0.50 mg/ml was lower than that of CD133- cells (Plt;0.05). ConclusionsCell population with CD133+ in the KATO-Ⅲ cell lines have b ability of cloning, better capability of proliferation and differentiation, as well as anticancer drug resistance to 5-FU. CD133 can be applied as one of surface markers for the detection to gastric cancer initiator cells.
Objective To investigate the clinical meanings of lymphangiogenesis, lymph vessel invasion (LVI) and lymph node (LN) micrometastasis in gastric cancer. Methods The expression of D2-40 in 68 patients with gastric cancer of primary lesion and the expressions of CK20 and (or) CKpan in 791 lymph nodes from 51 cases which were detected by immunohistochemical staining were analyzed, as well as their clinicopathologic profiles. The relationship of lymph vessel density (LVD), LVI and LN micrometastasis with LN metastasis and other clinicopathologic parameters was analyzed respectively. Results Positive rate of LVI with HE (LVI-HE) and D2-40 (LVI-IM) staining was respectively 66.2%(45/68) and 76.5%(52/68), P=0.118. The positive rate of LVI-IM was related to deeper tumor invasion (P=0.044), later stage of TNM (P=0.003) and LN metastasis (P=0.000). Average LVD of 68 cases was (18.19±7.44)/HP. The increment of LVD was significantly associated with LVI-HE positive status （P=0.040）, LVI-IM positive status （P=0.001）, venous invasion （P=0.037）, later stage of TNM （P=0.020） and LN metastasis （P=0.001）. The survival rate of the group sharing ≥15/HP of LVD was significantly lower than that in the group sharing ≤14/HP of LVD in early period of follow-up （P=0.032）. The incidence of nodal involvement in 51 patients was increased from 74.5%(38/51) by HE staining to 88.2%(45/51) by CK (CK20 or CKpan) immunostaining. The detection rate of metastasized LN was increased from 32.0%（253/791） by HE staining to 41.5%（328/791） by CK immunostaining (Plt;0.001). The significant difference of LN micrometastasis detection rate between CK20 (8.7%) and CKpan (12.3%) was also identified (P=0.003). The increased number of LN micrometastasis was related to larger diameter of tumor (P=0.001), higher LVI-HE positive rate (P=0.040), deeper invasion of tumor (P=0.018) and later stage of TNM (P=0.012). Both LN stage and TNM stage were changed according to the detection of LN micrometastasis: Seven patients of N0 should be recognized as N1 (N0→N1)， 6 as N1→N2, 1 as N2→N3. Four patients of stage Ⅰb should be recognized as stage Ⅱ (Ⅰb→Ⅱ)， 4 as Ⅱ→Ⅲa， 3 as Ⅲa→Ⅲb， 1 as Ⅲb→Ⅳ. Conclusion Detection of D2-40 and CK in diagnosis of LVI and LN micrometastasis is better than HE staining. The combined detection of CK20 and CKpan may be much easier to find out the LN with micrometastasis. Later stage of TNM the tumor is, more frequently the LN micrometastasis happens. The relationships of LVI-IM, LVD and LN micrometastasis with LN metastasis in gastric cancer has been demonstrated. Patients with higher LVD share a lower survival rate in early period of follow-up.
Objective To investigate the prognostic value of epithelial-mesenchymal transition （EMT） related proteins （Snail， E-cadherin， and N-cadherin） in gastric cancer and its relationship with tumor initiating cells （TICs） marker （CD133）. Methods The expressions of EMT-related proteins and CD133 protein in the gastric cancer tissues and normal gastric mucosa tissues adjacent to gastric cancer were detected by Western blot method. The relations between the expressions of EMT-related factors proteins and CD133 protein and the clinicopathologic characters were analyzed. The correlations between EMT-related factors and CD133 were analyzed by Spearman. The correlations between EMT-related factors expressions and CD133 expression and survival were analyzed by Kaplan-Meier method and Log-rank test. Results ① The protein expression levels of Snail， N-cadherin， and CD133 in the gastric cancer tissues were significantly higher than those in the normal gastric mucosa tissues adjacent to gastric cancer （Snail：0.599±0.114 versus 0.259±0.108， P=0.020；N-cadherin：0.754±0.154 versus 0.329±0.134， P=0.001；CD133：0.635±0.119 versus 0.485±0.116， P=0.029）， while the protein expression level of E-cadherin was lower than that in the normal gastric mucosa tissues adjacent to gastric cancer （0.378±0.123 versus 0.752±0.156， P=0.003）.② The expression levels of Snail and N-cadherin in the gastric cancer patients with vascular invasion， lymphatic vessel invasion，N3 lymph node metastasis， diameter more than 5 cm， and Ⅲ＋Ⅳ staging were significantly higher than those in the patients without vascular invasion， lymphatic vessel invasion， N0-N2 lymph node metastasis， diameter less than 5 cm， andⅠ＋Ⅱ staging（P＜0.05）， while E-cadherin protein expression was lower than that in the patients without vascular invasion， lymphatic vessel invasion， N0-N2 lymph nodes metastasis， andⅠ＋Ⅱstaging （P＜0.05）. The expression levels of CD133 in the gastric cancer patients with lymphatic vessel invasion， diameter more than 5 cm， and Ⅲ＋Ⅳ staging were significantly higher than those in the patients without lymphatic vessel invasion， diameter less than 5 cm， andⅠ＋Ⅱ staging （P＜0.05）. ③The Snail and N-cadherin protein expressions were significantly positive correlated with CD133 protein expression， respectively （rs=0.278， P=0.048；rs=0.406， P=0.003）， while E-cadherin protein expression was significantly negative correlated with CD133 protein expression （rs=-0.504， P=0.000）.④ The survival time in the patients with lower expressions of Snail， N-cadherin， and CD133 were significantly longer than those in the patients with higher expressions of Snail， N-cadherin， and CD133 （P＜0.05）. The combination of Snail， N-cadherin， E-cadherin， and CD133 could effectively predict survival. Conclusions There is a significant correlation between EMT and gastric cancer TICs， and which are correlated with aggressive clinicopathologic features of gastric cancer. The combination of Snail， E-cadherin， N-cadherin， and CD133 may be effectively predict the prognosis of gastric cancer patients.
ObjectiveTo investigate the expression of CD133 protein in primary lesions of gastric cancer and its clinical significance. MethodsThe expressions of CD133 protein in the primary lesion of tumor and normal gastric mucosa tissues confirmed by using histopathologic examination of 99 patients were detected by immunohistochemical staining. The correlation of CD133 protein expression with the clinicopathologic parameters and features after operation were analyzed. ResultsPositive cells of CD133 protein were localized in the gland parietal and cell membrane surface. The expression of CD133 protein in the cancer and normal gastric mucosa tissues were 29.29% (29/99) and zero, respectively (P=0.000). Expression of CD133 protein in tumor with diameter gt;5 cm was significantly higher than that in the tumor with diameter ≤5 cm (P=0.041). The expression of CD133 protein was correlated with TNM stage (P=0.044), lymph node metastasis (P=0.017), lymphatic vessel invasion (P=0.000), and vascular invasion (P=0.000). Logistic regression analysis revealed that invasion depth of tumor (P=0.011), lymph node metastasis (P=0.043), and TNM stage (P＝0.049) were independent risk factors for CD133 protein expression. Survival time of patients with positive expression of CD133 protein was significantly shorter than that negative expression of CD133 protein (P＝0.046). Cox proportial hazard regression model analysis demonstrated that lymph node metastasis (P=0.042), TNM stage (P=0.046), and positive expression of CD133 protein (P=0.046) were independent risk factors for patients survival. ConclusionThe CD133 protein expression in primary lesions is closely related with development, metastasis, and prognosis of gastric cancer.
ObjectiveTo investigate the expressions of contactin-1 (CNTN-1), vascular endothelial growth factor-C (VEGF-C), and its receptor VEGFR-3 (Flt-4) in primary gastric cancer and to explore the relevance among them and their correlation with clinicopathologic features of gastric cancer. MethodsThe VEGF-C, VEGFR-3, and CNTN-1 protein expressions of tumor tissues and normal gastric mucosa tissues in 68 patients with primary gastric cancer were analyzed by immunohistochemistry. The Flt-4-positive vessel density (FVD) and lymphatic vessel density (LVD) were also analyzed by VEGFR-3positive and D2-40-positive staining, respectively. ResultsThe positivity rate of VEGF-C, VEGFR-3, and CNTN-1 protein expression in the primary tumor was 57.4% (39/68), 60.3% (41/68), and 55.9% (38/68), respectively, which was significantly higher than that in the normal gastric mucosa tissues 〔20.6% (14/68), 23.5% (16/68), and 16.2% (11/68)〕， P=0.000. The expressions of VEGF-C, VEGFR-3, and CNTN-1 protein were significantly correlated with TNM stage, lymphatic vessel invasion, and lymph node metastasis (Plt;0.05). The expression of CNTN-1 protein was significantly correlated with VEGF-C (r=0.372, P=0.002) and VEGFR-3 protein expression (r=0.308, P=0.011). In tumor tissues of sixtyeight patients the FVD was (10.41±9.38)/HP, which was significantly lower than LVD 〔(18.19±7.44)/HP〕, P=0.000. Elevated FVD and LVD was significantly found in patients with tumor characterized by later TNM stage, severer lymphatic vessel invasion, and severer lymph node metastasis (Plt;0.05). The FVD of tumor was significantly correlated with VEGF-C (P=0.029) and CNTN-1 protein expression (P=0.003). The LVD of tumor was not significantly correlated with CNTN-1 (P=0.727), VEGF-C (P=0.173), and VEGFR-3 protein expression (P=0.924). The patients with positive expression of VEGF-C, VEGFR-3, and CNTN-1 protein showed poorer prognosis (Plt;0.05). ConclusionsElevated expression of CNTN-1 protein is observed in primary gastric cancer and correlated with VEGF-C and VEGFR-3 protein expression, indicating that combined detection has great value in prediction of invasive potential and prognosis. VEGF-C-mediated CNTN-1 overexpression may promote lymphatic invasion via lymphangiogenesis pathway in patients with gastric cancer.
Objective To investigate the expression of transcription factor e2f-1 in the different development stages of gastric cancer, the relationships between clinicopathologic characteristics and e2f-1 expression status, as well as its influences on the prognosis. Methods The operative samples from primary lesion of 121 patients who underwent radical resection for gastric cancer were detected by SABC immunohistochemical staining. The relationships of e2f-1 expression with clinicopathologic characteristics and with the prognosis were observed by univariate, multivariate and relative analyses. Results The total positive expression rate of e2f-1 in all patients was 38.8% （47/121）. With the advancement of gastric cancer, the level of e2f-1 expression in TNMⅠ-Ⅳ stage was gradually decreased (r=－0.320， Plt;0.05)： Ⅰa stage with 62.5% （10/16）， Ⅰb with 47.1% （8/17）， Ⅱwith 55.0% （11/20）， Ⅲa with 40.0% （8/20）， Ⅲb with 27.3% （6/22）， Ⅳ with 15.4% （4/26）. The expression of e2f-1 was significantly negative correlated with tumor diameter, depth of infiltration, lymph node metastasis ratio, and N stage （Plt;0.05）. The multivariate analysis revealed that either histology type, or survival time was respectively an independent factor for e2f-1 expression （Plt;0.05）. Log-Rank test showed the relative factors to survival included N stage, tumor diameter, tumor position, lymph node metastasis ratio, depth of infiltration, and TNM stage （Plt;0.05）. Cox survival analysis found that both of later N stage and e2f-1 higher expression were independent prognostic factors （Plt;0.05）. The higher e2f-1 expression was related to a poor survival in TNM stageⅠand Ⅱ patients （r=－0.304, Plt;0.05）, the prognosis of patients with e2f-1 positive expression was worse than that of patients with negative expression （χ2=13.437， Plt;0.05）, and there was no statistic relationship between the expression of e2f-1 and prognosis in stage Ⅲ and Ⅳ patients （Pgt;0.05）. Conclusions e2f-1, as a useful marker, seems to be an indication for the malignant behavior in relatively earlier gastric cancer, in which the e2f-1 positive expression shares a significantly poor survival. And the lower expression of e2f-1 has been identified in later advanced gastric cancer, the more malignances in advanced gastric cancer might associate with a lower expression of e2f-1.