ICAM-1 (intercellular adhesion molecule-1) gene transfection inhibits lymph node metastasis by human gastric cancer cells.

Lymph node metastasis is one of the prognostic factors in gastric cancer. We have previously reported that decreased intercellular adhesion molecule-1 (ICAM-1) expression on cancer cells is associated with lymph node metastasis using a gastric cancer cell. In this study, we transfected ICAM-1 gene into a gastric cancer cell line, 2MLN, and analyzed the effect on lymph node metastasis in vitro and in vivo. A significantly greater amount of peripheral blood mononuclear cells (PBMC) adhered to ICAM-1 transfected 2MLN cells, 2MLN / ICAM cells, than to 2MLN / Vector cells. The lysis of 2MLN / ICAM cells by PBMC was significantly increased compared with that of 2MLN / Vector cells. The tumor growth rate of 2MLN / ICAM cells was significantly decreased in vivo. Lymph node metastases caused by 2MLN / ICAM cells were recognized as being fewer in number and smaller, while many lymph node metastases were caused by 2MLN cells. Histologic findings showed that leukocytes were heavily infiltrated in both the 2MLN / ICAM tumors and metastatic lesions, while only a few leukocytes were observed in the lesions associated with 2MLN cells. The above findings indicate that ICAM-1 gene transduction could prove to be an effective gene therapy for lymph node metastasis of gastric cancer.     

Up regulation of ICAM-1 gene expression inhibits tumour growth and liver metastasis in colorectal carcinoma

We have previously reported that decreased intercellular adhesion molecule-1 (ICAM-1) expression in cancer cells is associated with liver metastasis of colorectal cancer. In this study, we have investigated the effect of ICAM-1 gene transfection into the human colorectal cancer cell line LM-H3 on cell adhesiveness and cytotoxicity of peripheral blood mononuclear cells (PBMC) to cancer cells. Furthermore, we have investigated the effects of this gene transfer on subcutaneous tumour and liver metastases of LM-H3 in nude mice. More PBMC adhered to ICAM-1 transfected LM-H3 cells, LM-H3/ICAM-1, than to non-transfected LM-H3 cells and control LM-H3/Vector. Lysis of LM-H3/ICAM-1 cells by PBMC was significantly increased compared with LM-H3/Vector. Liver metastases with LM-H3/ICAM-1 cells were fewer in number and smaller than metastases with LM-H3/Vector. Intra-tumoural injection of ICAM-1 adenoviral vector significantly inhibited the growth of subcutaneous LM-H3 tumour. In conclusion ICAM-1 gene transfection using adenovirus vector might be an effective therapy for liver metastasis of colorectal carcinoma.     

Decrease in ICAM-1 expression on gastric cancer cells is correlated with lymph node metastasis

Background. Lymph node metastasis is a frequent type of metastasis in patients with gastric cancer. The mechanisms responsible for this type of metastasis, however, are not clearly understood. We hypothesize that the immunosurveillance system between cancer cells and lymphocytes may be associated with the lymph node metastatic process. In this study, we examined the correlation between lymph node metastasis and intercellular adhesion molecule-1 (ICAM-1), which mediates the immunosurveillance system between tumor cells and cytotoxic lymphocytes, in gastric cancer. Methods. One hundred and forty-three specimens resected from patients with gastric cancer were investigated by staining with a monoclonal antibody against ICAM-1. We studied the correlation between the expression of ICAM-1 and various clinicopathologic factors, as well as infiltration of tumor-infiltrating lymphocytes (TILs). Results. ICAM-1 expression on gastric cancer cells was significantly decreased in patients with lymph node metastasis. The infiltration of TILs was associated with ICAM-1 expression level. The prognosis of patients with ICAM-1-negative tumors was poorer than the prognosis of those with ICAM-1-positive tumors. Conclusions. These findings suggest that ICAM-1 expression on cancer cells is closely associated with lymph node metastasis in gastric cancer, under the influence of the host immunosurveillance system. Received on Aug. 20, 1999; accepted on Jan. 5, 2000     

Effect of a matrix metalloproteinase inhibitor on a lymph node metastatic model of gastric cancer cells passaged by orthotopic implantation

Lymph node metastasis is the most frequent type of tumor recurrence and is known to be one reason for the poor prognosis in patients with digestive cancer. However, the mechanisms of lymph node metastasis are not clearly understood and a metastatic model will be useful for elucidating factors associated with lymph node metastases. In this study, we investigated the effect of R-94138, a matrix metalloproteinase (MMP) inhibitor, on the lymphnodal metastatic ability of gastric cancer cells using an in vivo orthotopic implantation model in nude mice. Injection of a gastric cancer cell line, MKN-45, into the gastric wall resulted in lymph node metastasis 8 weeks after inoculation. The number of lymph node metastases and the amount of body weight loss significantly decreased by intraperitoneal administration of R-94138. Histologically, lymphatic invasion of cancer cells was found in primary gastric tumors of control mice with lymph node metastasis. However, no lymphatic invasion was observed in the gastric wall of R-94138-treated mice without lymph node metastasis. These findings suggested that the MMP inhibitor, R-94138, could be used in adjunctive therapy for lymph node metastasis in gastric carcinoma.     

CD80 gene therapy for lymph node involvement by gastric carcinoma

The co-stimulatory molecule, CD80 (B7-1), is a ligand of CD28 and plays a key role in the induction of cell-mediated immune responses. Many tumors, including gastric cancer, have decreased expression of CD80 which leads to a failure of immune recognition. Lymph node spread is a factor of poor prognosis in gastric cancer. In this study, we transfected the CD80 gene by an adenovirus vector into a human gastric cancer cell line, OCUM-2MLN, and analyzed the effect on lymph node disease in vitro and in vivo. After transfection of CD80 in vitro, the adhesive ability of cancer cells for peripheral blood mononuclear cells and their cytotoxicity showed significant regression (p<0.01). Intratumoral injection of AdCD80 caused significantly growth of subcutaneous tumors. In vivo lymph node spread was suppressed by injection of AdCD80 into gastric tumors. Histopathologic findings revealed CD80-positive cells around the tumor. These results suggest that CD80 gene transfer into cancer cells using an adenovirus vector might be a promising approach for in vivo cancer therapy.     

Combination effect of a TGF‐β receptor kinase inhibitor with 5‐FU analog S1 on lymph node metastasis of scirrhous gastric cancer in mice

Transforming growth factor‐β (TGF‐β) signals are closely associated with the distant metastases of gastric cancer. The aim of this study was to clarify the effect of a TGF‐β receptor I (TβR‐I) phosphorylation inhibitor, Ki26894, in combination with anticancer drugs, on the lymph node (LN) metastasis of scirrhous gastric cancer. A novel TβR‐I kinase inhibitor, Ki26894, inhibits the phosphorylation of Smad2 at the ATP binding site of TβR‐I. S1 is a 5‐fluorouracil analog. The human scirrhous gastric cancer cell line OCUM‐2MLN and the human gastric fibroblasts NF‐33 were used. OCUM‐2MLM cells in the upper well and NF‐33 cells in the lower well were co‐incubated with or without Ki26894. The proliferation of OCUM‐2MLN cells was significantly stimulated by co‐culture with NF‐33 cells. Ki26894 significantly suppressed the growth interactions between OCUM‐2MLN cells and NF‐33 cells. Gastric cancer models established by orthotopic inoculation of OCUM‐2MLN cells showed diffusely infiltrating gastric adenocarcinoma accompanied by LN metastases. We divided these mice into four groups, (control vehicle, Ki26894, S1, Ki26894 plus S1), and examined the effect of Ki26894 and/or S1 on phosphorylation of Smad2, tumor size, LN metastases, and lymphatic involvements. Ki26894 inhibited the Smad2 phosphorylation of cancer cells and decreased the extent of lymphatic involvement, compared with the control or S1 only group. The Ki26894 plus S1 administration group significantly suppressed tumor growth and decreased LN metastasis more effectively than either alone. These findings suggested that the TβR‐I kinase inhibitor with S1 is useful for the treatment of scirrhous gastric carcinoma with LN metastasis. (Cancer Sci 2010)     

Galectin-1蛋白及孕激素受体与胃癌腹膜转移的关系

目的：探讨胃癌组织中半乳糖结合蛋白（Galectin-1）及孕激素受体（PR）表达与胃癌腹膜转移的关系。方法：应用即用型孕激素测定试剂盒及鼠抗人Galectin-1单克隆抗体和免疫组织化学SP染色法检测40例同期伴腹膜转移胃癌患者的癌旁正常胃黏膜、胃癌原发灶、胃癌腹膜转移灶及淋巴结转移灶中Galectin-1蛋白及PR的表达。结果：正常胃黏膜中Galectin-1及PR的表达与胃癌原发灶、腹膜转移灶及淋巴结转移灶之间差异均具统计学意义（P〈0．05）。胃癌原发灶、腹膜转移灶及淋巴结转移灶三者间的Galectin-1及PR表达差异无统计学意义（P〉0．05）。结论：胃癌病灶中的Galectin-1及PR表达可能成为判断胃癌患者术前是否有腹膜转移及预后的生物学指标,并可为胃癌的内分泌治疗提供依据。     

ICAM-2 gene therapy for peritoneal dissemination of scirrhous gastric carcinoma.

PURPOSE: Human scirrhous gastric carcinoma develops peritoneal dissemination with high frequency, and the prognosis of patients with peritoneal metastasis is poor. There have been few reports of an immunogene therapy for peritoneal dissemination. Intercellular adhesion molecule (ICAM)-2 is a second ligand of leukocyte function-associated antigen-1, which functions as a costimulatory molecule for effector cells. In the present study, we examined whether ICAM-2 transfection using adenovirus vector is effective gene therapy for peritoneal metastasis of gastric cancer. EXPERIMENTAL DESIGN: We constructed an adenovirus vector, AdICAM-2, that encodes the full-length human ICAM-2 gene under control of the cytomegalovirus promoter. This vector expresses high levels of ICAM-2 on the human gastric cancer cell line OCUM-2MD3, which has high peritoneal metastatic ability in nude mice. We investigated the antitumor effects of gene transfer of ICAM-2 using the adenovirus vector AdICAM-2 in vitro and in vivo. RESULTS: ICAM-2 expressed on OCUM-2MD3 cells by AdICAM-2 demonstrated significantly high adhesiveness to and cytotoxicity against peripheral blood mononuclear cells in vitro compared with the control adenovirus vector AdlacZ. Intratumoral injection of AdICAM-2 significantly inhibited the growth of s.c. tumor. Mice with peritoneal metastasis survived for a significantly longer time after AdICAM-2 injection, compared with injection of AdlacZ. Histopathological findings revealed that many natural killer cells infiltrated the peritoneal metastatic lesions after AdICAM-2 injection. CONCLUSIONS: These findings suggest that transduction of ICAM-2 into cancer cells enhances the adhesion and activation of natural killer cells, resulting in a reduction of peritoneal metastasis. ICAM-2 transfection using adenovirus vector might be an effective form of gene therapy for peritoneal metastasis of gastric cancer.     

Suppression of lymph node and lung metastases of endometrial cancer by muscle‐mediated expression of soluble vascular endothelial growth factor receptor‐3

Lymph node metastasis is the most important prognostic factor of endometrial cancer. However, effective therapy has not been established against lymph node metastasis. In this study, we explored the efficacy of gene therapy targeting lymph node metastasis of endometrial cancer by suppressing the action of vascular endothelial growth factor (VEGF)‐C through soluble VEGF receptor‐3 (sVEGFR‐3) expression. For this purpose, we first conducted a model experiment by introducing sVEGFR‐3 cDNA into an endometrial cancer cell line HEC1A and established HEC1A/sVEGFR‐3 cell line with high sVEGFR‐3 expression. The conditioned medium of HEC1A/sVEGFR‐3 cells inhibited lymphatic endothelial cell growth in vitro, and sVEGFR‐3 expression in HEC1A cells suppressed in vivo lymph node and lung metastases without inhibiting the growth of a subcutaneously inoculated tumor. To validate the therapeutic efficacy, adeno‐associated virus vectors encoding sVEGFR‐3 were injected into the skeletal muscle of mice with lymph node metastasis. Lymph node and lung metastases of HEC1A cells were completely suppressed by the muscle‐mediated expression of sVEGFR‐3 using adeno‐associated virus vectors. These results suggest the possibility of gene therapy against lymph node and lung metastases of endometrial cancer by using muscle‐mediated expression of sVEGFR‐3.     

Development of a mouse model for lymph node metastasis with endometrial cancer

Controlling lymph node metastasis is currently a key issue in cancer therapy. Lymph node metastasis is one of the most important prognostic factors in various types of cancers, including endometrial cancer. Vascular endothelial growth factor-C (VEGF-C) plays a crucial role in lymphangiogenesis, and is implicated to play an important role in lymph node metastasis. To evaluate the role of VEGF-C in lymph node metastasis, we developed an animal model by using an endometrial cancer cell line, HEC1A. This cell line is not invasive by nature and secretes moderate amounts of VEGF-C; intrauterine injection of HEC1A cells into Balb/c nude mice resulted in uterine cancer with lymph node metastasis after 8 weeks. To analyze the contribution of VEGF-C to lymph node metastasis, its corresponding gene was stably introduced into HEC1A cells (HEC1A/VEGF-C), which then produced more than 10 times the amount of VEGF-C. The number of lymph node metastases was significantly higher in HEC1A/VEGF-C cells than in HEC1A cells (3.2 vs 1.1 nodes/animal, respectively). Augmented lymphangiogenesis was observed within tumors when HEC1A/VEGF-C cells were inoculated. These results indicate that VEGF-C plays a critical role in lymph node metastasis, in addition to serving as a platform to test the efficacy of various therapeutic modalities against lymph node metastasis.     

ICAM-1 expression and the soluble ICAM-1 level for evaluating the metastatic potential of gastric cancer

ICAM-1 plays an important role in cell-cell and cell-extracellular matrix interactions, especially tumor invasion and cytotoxicity of lymphocytes. In the present study, the relationship between metastasis of gastric cancer and ICAM-1 expression by cancer cells or the serum level of s-ICAM-1 was (s-ICAM-1) was examined. ICAM-1 was detected by immunohistochemic staining in 49.0% of 108 patients with gastric cancer. The ICAM-1 expression rate was higher at a more advanced stage, based on lymph node metastasis, being 46.9% in node-negative and 56.1% in node-positive cases. In patients with liver metastasis, the rate was 90.9%, while it was 43.3% in patients without liver metastasis (p < 0.05). The serum s-ICAM-1 level was 262.1 ng/ml (median 205.5, range 176.0-271.0) in healthy subjects and 391.5 ng/ml (median 317.5, range 148.7-1,768.0) in gastric cancer patients (p < 0.001). The serum s-ICAM-1 level was significantly higher in patients with liver metastasis than in patients without liver metastasis (p < 0.0001). In addition, positive ICAM-1 expression cases had significantly higher s-ICAM-1 levels than negative ones, 408.9 +/- 188.4 and 308.1 +/- 88.1 ng/ml, respectively. These results suggested that ICAM-1 was overexpressed in cancer cells and released as s-ICAM-1, which would promote hematogenous metastasis by suppressing local anticancer immunity.     

Establishment and characterization of a human gastric carcinoma cell line that is highly metastatic to lymph nodes

The actual mechanisms by which carcinoma cells metastasize to lymph nodes are still unclear, and there is a need to establish in vivo experimental models suitable for the investigation of lymph node metastasis. For the purpose, we established a highly lymph node-metastasizing line, designated AZL5G, derived from a human gastric cancer cell line, AZ521, which had low capacity for lymph node metastasis. AZL5G cells transplanted orthotopically in the nude mouse stomach metastasize predominantly to the regional lymph nodes, showing little potential for hematogenous metastasis. AZL5G tumors developing in the stomach and regional lymph nodes showed poorly differentiated adenocarcinoma with medullary growth, and their histologic appearance strongly resembled that of parental AZ521. The growth activities in vitro of low-metastatic AZ521 and high-metastatic AZL5G were almost the same, but the tumorigenicity in vivo of AZL5G was significantly higher than that of AZ521. AZL5G cells also showed clearly higher abilities of cell locomotion and adhesion to type IV collagen and fibronectin in vitro as compared with AZ521 cells. Flow cytometric analysis demonstrated that the expression of integrin beta1 subfamily except for alpha6 integrin was generally increased in AZL5G cells than in AZ521 cells. Especially, the expression of alpha1 and alpha2 integrins in AZL5G cells was clearly higher than in AZ521, while alpha(v)beta3 integrin, E-cadherin, ICAM-1 and CD44H were not expressed by either cell line. The cell adhesion blocking assay showed that DGEA-containing peptide, which is composed of alpha2 integrin recognition sequence, significantly reduced the adhesiveness of AZL5G cells to type IV collagen as well as to type I collagen and laminin. Furthermore, the administration of anti-alpha2 integrin mAb or DGEA peptide in AZL5G-transplanted nude mice produced a significant reduction in the number of lymph node metastases. These data suggest that the up-regulation of alpha2 integrin expression by gastric cancer cells may play a critical role in the process of lymph node metastasis through the increased adhesiveness to type IV collagen. In conclusion, we established a gastric cancer cell line, AZL5G, with a highly metastatic potential to lymph nodes. This well-characterized line and its in vivo experimental model should be useful for investigation of the mechanisms of lymph node metastasis and for establishment of a new therapeutic approach for human gastric cancer.     

Clinicopathological Characteristics as Predictive Factrs for Lymph Node Metastasis in Submucosal Gastric Cancer

OBJECTIVE To identify clinicopathological characteristics as predictive factors for lymph node metastasis in submucosal gastric cancer, and in addi- tion to establish objective criteria as indications for endoscopic submucosal dissection (ESD). METHODS Data from 130 patients with submucosal gastric cancer were collected, and the relationship between their clinicopathological characteris- tics and the presence of lymph node metastasis was retrospectively analyzed by multivariate analysis. RESULTS In the multivariate logistic regression model, a tumor size of 2 cm or more and an undifferentiated histologic type were found to be inde- pendent risk clinicopathological characteristics for lymph node metastasis. Among 130 patients with submucosal carcinoma, no lymph node metastases were observed in 17 patients who showed neither of the two risk clinicopath- ological characteristics. Lymph node metastasis occurred in 61.1% (22/36) of the patients who had both risk clinicopathological characteristics. CONCLUSION A tumor size of 2 cm or more and an undifferentiated histologic type were significantly and independently related to lymph node metastasis in submucosal gastric cancer. It is rational for the paitients with neither of these two independent risk clinicopathological characteristics to undergo an ESD.     

淋巴管癌栓和淋巴结转移对胃癌术后生存率的影响

目的：探讨淋巴管癌栓与胃癌临床病理特点的关系。方法：比较淋巴管栓和淋巴结转移对胃癌预后的影响。结果：１）浸润深度及生长方式影响淋巴管癌栓的发生；２）只有在元淋巴结转移的条件下，淋这癌栓才会影响胃癌患者的预后。结论：淋巴结转移对一影响明显高于淋巴管癌栓。     

直肠癌淋巴结转移规律的临床研究

目的探讨直肠癌区域淋巴结转移规律及术中判断淋巴结转移的准确性。方法回顾性分析180例根治性手术后的直肠癌临床资料,分析淋巴结转移情况。结果患者的性别、年龄和肿瘤的大小与淋巴结转移无明显关系,肿瘤浸润肠壁的深度及肿瘤的分化程度与淋巴结转移有关。T3～T4时,其淋巴结阳性率和转移度显著高于T1～T2者(P＜0.05),低分化肿瘤的淋巴结阳性率和转移度明显高于高分化肿瘤(P＜0.05)。51例术中发现淋巴结肿大,129例未发现淋巴结肿大,术后病理证实分别有31例和77例淋巴结转移阳性。结论直肠癌淋巴结转移与肿瘤浸润肠壁的深度及分化程度有关,手术中淋巴结是否转移不准确,因此应彻底切除直肠区域淋巴结。     

Predictive value of cathepsin D and Ki-67 expression at the deepest penetration site for lymph node metastases in gastric cancer

To search for reliable predictors for lymph node metastasis, we immunohistochemically analyzed surgically resected gastric cancer specimens that showed invasion of submucosa (sm) and muscularis propria (mp) of the tumor. The analysis investigated cathepsin D and Ki-67 expression in 136 specimens that were divided into an sm1/sm2 group and an sm3/mp group. In sm1/sm2 group, the incidence of lymph node metastases was significantly higher in tumors with high Ki-67 labeling index (LI) (44%) than in those with low Ki-67 LI (0%). In sm3/mp group, the incidence of lymph node metastases was significantly higher in cathepsin D-positive (56%) and high Ki-67 LI tumors (64%) than in cathepsin D-negative (33%) and low Ki-67 LI (33%). Combined analysis of cathesin D expression and Ki-67 LI correlated strongly with lymph node metastases. No lesions with cathepsin D-negative expression and low Ki-67 LI had lymph node metastases in either group. Cathepsin D and Ki-67 expression may be useful predictors for lymph node metastases in gastric cancer with sm and mp invasion. As predictors, they can identify lesions without lymph node metastases and indicate lesions not needing additional treatment after endoscopic mucosal resection and laparoscopic gastrectomy.     

早期胃癌淋巴结转移因素分析

目的探讨影响早期胃癌淋巴结转移的因素。方法对74例术后早期胃癌患者的资料,对各临床病理指标与淋巴结转移的关系进行分析,以确定淋巴结转移的危险因素。结果早期胃癌患者的淋巴结转移率为14.9%（11/74）。单因素分析显示黏膜下癌的淋巴结转移率（27.6%）明显高于黏膜内癌（6.7%）（P=0.020）;未分化型癌的淋巴结转移率（27.6%）明显高于分化型（6.8%）（P=0.042）;肿瘤最大径≤2 cm、〉2-4 cm、〉4 cm 3组间淋巴结转移率有统计学意义（χ2=6.549,P=0.038）。采用Log istic回归进行的多因素分析显示,肿瘤最大径（OR=2.688,P=0.047）和浸润深度（OR=4.508,P=0.044）是影响早期胃癌淋巴结转移的独立危险因素。结论早期胃癌淋巴结转移与肿瘤最大径和浸润深度密切相关,这可为手术方案的选择提供参考。     

MSCT对T1和T2期非小细胞肺癌患者纵隔淋巴结转移的诊断意义

  目的   评估胸部多层CT扫描（MSCT）对T₁和T₂期非小细胞肺癌（NSCLC）纵隔淋巴结转移的指导意义。   方法   选择2004年3月至2012年3月T₁和T₂期NSCLC患者32例，依据病理结果分析术前MSCT对纵隔淋巴结的判断。   结果   以淋巴结短径≥10 mm MSCT评价纵隔淋巴转移的敏感性和特异性分别为82.4%和92.4%；淋巴结大小、原发肿瘤位置及脏胸膜侵犯对纵隔淋巴转移的预测差异均有统计学意义（P < 0.05）。   结论   淋巴结大小可作为评估NSCLC患者纵膈淋巴结转移的依据，原发于右肺的肿瘤及肿瘤伴有脏层胸膜侵犯具有较高的纵隔淋巴结转移风险。      

早期胃癌组织中上皮钙粘蛋白表达与淋巴结微转移及临床病理的关系

背景与目的:早期胃癌淋巴结微转移问题日益受到关注,胞浆角蛋白(cytokeratin,CK)染色是识别上皮源性恶性肿瘤细胞的重要方法,本研究拟探讨早期胃癌原发灶上皮钙粘蛋白(epithelial cadherin,E-cad)的表达情况与淋巴结内出现微转移之间的关系及临床意义.方法:用免疫组织化学染色的方法对162例早期胃癌患者的4 522枚淋巴结进行苏木精-伊红(HE)和胞浆角蛋白(cytokeratin,CK)染色,并对其中135例患者的原发灶切片进行E-cad染色,结合临床病理资料和随访结果进行分析.结果:HE染色发现的淋巴结转移率为6.8%(11/162),而CK染色发现的淋巴结转移率为26.5%(43/162),二者差异有统计学意义(P＜0.001).在151例HE染色未见淋巴结转移的患者中通过CK染色发现了32例(21.2%)有淋巴结微转移,且淋巴结微转移多见于原发灶直径大于1.0 cm,组织分化不良,肿瘤浸润较深的(如浸及粘膜下层),淋巴管和血管受累,以及E-cad低表达标本(P＜0.05).原发灶E-cad的低表达率为57.0%(77/135),与淋巴结出现微转移有密切关系,有淋巴结微转移患者的5年生存率比没有微转移者明显低(P＜0.01).结论:肿瘤直径大于1.0 cm,组织分化不良,较深的浸润,淋巴管或血管受累,以及E-cad低表达是早期胃癌患者出现淋巴结转移的高危因素.