Superselective Drug Delivery Using Doxorubicin-Encapsulated Liposomes and Ultrasound in a Mouse Model of Lung Metastasis Activation

Conventional treatment of lymph node metastasis involves dissection of the tumor and regional lymph nodes, but this may cause activation of latent metastatic tumor cells. However, there are few reports on animal models regarding the activation of latent metastatic tumor cells and effective methods of treating activated tumor cells. Here, we report the use of a superselective drug delivery system in a mouse model of lung metastasis in which activated tumor cells are treated with doxorubicin-encapsulated liposomes (DOX-LP) and ultrasound. The axillary lymph node was injected with DOX-LP and exposed to ultrasound so that the released DOX would be delivered from the axillary lymph node to the metastatic lung via the subclavian vein, heart and pulmonary artery. The size of the DOX-LP was optimized to a diameter of 460?nm using indocyanine green-encapsulated liposomes, and the ultrasound intensity was 0.5 W/cm². We found that compared with DOX or DOX-LP alone, the superselective drug delivery system was effective in the treatment of metastasis in both the lung and axillary lymph node. We anticipate that this superselective drug delivery system will be a starting point for the development of new techniques for treating lung metastasis in the clinical setting. Furthermore, the superselective drug delivery system may be used to screen novel drugs for the treatment of lung cancer and investigate the mechanisms of tumor cell activation after resection of a primary tumor or lymph nodes.     

Vitamin D3 treatment to diminish the levels of immune suppressive CD34+ cells increases the effectiveness of adoptive immunotherapy

Tumor growth can increase the number of immature bone marrow-derived CD34+ cells that exhibit natural suppressor (NS) activity toward T-cell function. Using a metastatic Lewis lung carcinoma (LLC-LN7) tumor model, these CD34+ NS cells were shown to be present within the s.c. primary tumor tissue, but their levels declined after treatment with the inducer of myeloid cell differentiation, vitamin D3. Therefore, studies determined whether vitamin D3 treatment to diminish the CD34+ NS cell levels in LLC-LN7-bearing mice would enhance (a) intratumoral immune reactivity and (b) the antitumor activity of adoptive therapy consisting of tumor-reactive lymph node cells. The results showed that vitamin D3 treatment alone increased the intratumoral CD8+ cell content and the activity of the intratumoral infiltrate, as detected by production of interferon-gamma and expression of the p55 IL-2 receptor. Although vitamin D3 treatment had no effect on the size of the primary tumor, it lessened the extent of tumor metastasis. Treating mice with the combination of vitamin D3 and adoptive immunotherapy significantly reduced metastasis in mice with established tumors, and reduced both metastasis and locoregional recurrence after surgical excision of the primary tumor. These studies demonstrate that vitamin D3 treatment increases intratumoral T-cell immune reactivity, and that coupling vitamin D3 treatment to diminish levels of CD34+ NS cells with adoptive immunotherapy enhances the effectiveness of the adoptively transferred tumor-reactive lymph node cells at limiting both metastasis and locoregional tumor recurrence.     

Aspirin reduces lung cancer metastasis to regional lymph nodes

Background

Lung cancer is the main cause of cancer-related death worldwide. The high mortality is probably attributable to early metastasis; however, the mechanism underlying metastasis to regional lymph nodes is still unknown. Cyclooxygenase (COX)-derived prostaglandin E₂ (PGE₂) induces tumor growth and metastasis and is associated with a poor prognosis. The present study investigated the effect of an authentic COX inhibitor, aspirin, on regional lymph node metastasis during the development of lung cancer in mice.

Methods

An orthotopic intrapulmonary implantation model based on male C57BL/6 (6–8-weeks-old) mice was used. The lungs were injected with a solution containing Lewis lung carcinoma (LLC) cells overexpressing green fluorescent protein (GFP) and BD Matrigel^®. The effect of aspirin on mediastinal lymph node metastasis of LCC cells from the primary injection sites was then examined.

Results

The implantation process took approximately 30 s per mouse and operative mortality was 10%. Single pulmonary nodules developed at the implanted site in 95% of animals, and regional mediastinal lymph node metastasis was observed at 14 days post-LLC-GFP cell injection in all mice that formed a primary lung tumor. The mean survival time of mice injected with LLC-GFP cells was 15 ± 3 days (range, 12–22 days). Histopathological analysis revealed that no metastatic tumors developed in the regional mediastinal lymph nodes by Day 10–12 post-LLC-GFP cell injection and no metastasis to distant organs or distant lymph nodes was observed by Day 21 post-injection. Oral administration of aspirin (100 mg/kg, twice a day) after LLC-GFP cell injection inhibited metastasis to the regional lymph nodes, with no significant suppression of primary tumor growth in the lungs. Aspirin treatment led to a significant reduction in mortality (P < 0.0001).

Conclusions

The present lymph node metastasis model is useful for evaluating the efficacy of agents that inhibit tumor metastasis to the regional lymph nodes. Aspirin reduced the metastasis of LLC-GFP cells injection to the regional lymph nodes, with a significant reduction in mortality. These findings suggested that COX inhibitors have potential for preventing lymph node metastasis.     

非小细胞肺癌淋巴结转移规律的临床病理学特征分析

目的探讨非小细胞肺癌( NSCLC)患者淋巴结转移规律的临床病理学特征。 方法对80例NSCLC患者施行患侧肺手术切除并行广泛肺门、肺叶间及纵隔淋巴结清扫术,共清扫602组淋巴结,分析NSCLC的淋巴结转移规律与临床病理特征的关系,并结合T分期、病理学类型、分化程度及原发部位进行统计分析。 结果在共清扫的602组淋巴结中,单纯N1淋巴结转移率为18.8%,N2淋巴结转移率30.4%;原发性NSCLCT分期T1、T2、T3、T4间淋巴结转移率差异有统计学意义(χ²＝50.702,P＝0.000);跳跃式转移占N2转移的31.8%。 结论NSCLC的淋巴结转移与T分期有关,具有较多的跳跃性纵隔淋巴结转移发生,肿瘤部位及肺癌的病理学类型与淋巴结的转移无明显关系。     

Matrix metalloproteinase inhibitor MMI-166 inhibits lymphogenous metastasis in an orthotopically implanted model of lung cancer

Matrix metalloproteinases (MMP) are considered to be critically involved in tumor invasion and the metastasis of various cancers. MMI-166 is a selective inhibitor of matrix metalloproteinase (MMP-2, MMP-9, and MMP-14). The purpose of this study was to evaluate the effects of MMI-166 on both the growth of the implanted tumor and the lymph node metastasis of the mediastinum and prolonging the life span, using an orthotopic implantation model of the Ma44-3 cancer cell line. We examined the anti-invasive effect of MMI-166 in lung cancer cell lines using an in vitro invasion assay. Next, we examined the anticancer effect of MMI-166 in vivo. MMI-166 (200 mg/kg body weight) or a vehicle was administered orally to the orthotopically implanted lung cancer model. MMI-166 dose-dependently inhibited the invasion of cancer cell lines with expressions of MMP-2 and/or MMP-9 in vitro. In vivo, MMI-166 significantly inhibited mediastinal lymph node metastasis in this orthotopic model (weight of the mediastinum: control, 0.089 +/- 0.009 versus MMI-166, 0.069 +/- 0.008 mg; P = 0.005; metastatic area: control, 93,495 +/- 55,747 versus MMI-166, 22,747 +/- 17,478 pixels; P = 0.045). MMI-166 prolonged the life span by 6 days in median survival time in the orthotopically implanted model (P = 0.039). These results showed that MMI-166 could possibly inhibit lymph node metastasis and prolong the life span in lung cancer patients.     

Natural cytostatic cells have a broader range of antitumor activity than natural killer cells

Natural cytotoxic activity by Thy-1 negative, asialo-GM-1 positive and column-nonadherent spleen cells was detected against YAC-1 target cells but not against Meth-A and EL-4 target cells as reported by many investigators. In the present study, on the other hand, natural cytostatic activity was detected in Thy-1 negative, asialo-GM-1 negative and column-nonadherent cells and this activity was effective against YAC-1, Meth-A and EL-4 target cells. Such a cytostatic activity was not modified by the functional levels of T cells in the donor mice of effector cells. The cytostatic activity against Meth-A and EL-4 target cells was detected not only in an allogeneic system but also in a syngeneic system. The cytostatic activity was not depressed in a tumor-bearing state in contrast to the NK activity. The cytostatic activity as well as the NK activity was detected in spleen and peritoneal cells but not in lymph node cells of the various sources. The spleen and peritoneal cavity may play a role distinct from that of lymph nodes in the resistance against tumor development.     

VCPmRNA在胃癌中的表达及意义

目的研究VCPmRNA表达与胃癌及淋巴结转移的关系，探讨其术前预测淋巴结转移危险度及预后的意义。方法采用RT—PCR、聚丙烯酰胺凝胶电泳EB染色法，检测42例胃癌组织、癌旁正常胃组织、淋巴结组织中VCPmRNA的表达。结果VCPmRNA在上述组织细胞中均有表达，胃癌组织和淋巴结组织明显高于癌旁正常胃组织（P〈0.001），胃癌组织VCPmRNA表达丰度随转移淋巴结枚数的增多而增高，转移淋巴结数量≥16枚者高于〈16枚者（P〈0.05）。结论胃癌组织VCPmRNA过度表达的患者淋巴结转移范围及程度相对较重，癌细胞侵袭力较强，宜行扩大根治术；术前RT—PCR检测胃癌组织VCPmRNA表达有助于确定治疗方案及判断预后。     

胸腔镜手术与开胸手术在胸段食管癌患者淋巴结清扫中的对比

目的探讨胸腔镜手术与开胸手术下胸段食管癌患者的淋巴结清扫状况。方法选取2012年8月-2015年6月在该院接受胸腔镜手术治疗的胸段食管癌患者16例(微创组),选取同期经传统开胸手术的相同病理分期的胸段食管癌患者17例(传统组),比较两组患者清扫淋巴结数、阳性率及淋巴结转移情况。结果微创组16例,清扫淋巴结总数228枚,均数(14.27±5.61)枚,阳性率为12.50%;传统组17例,清扫淋巴结总数241枚,均数(16.20±6.24)枚,阳性率为11.76%。两组不同病理分期患者淋巴结清扫数差异无统计学意义(P0.05)。微创组转移淋巴结例数7例,转移淋巴结45枚,转移率为43.75%,转移度为19.74%;传统组转移淋巴结例数8例,转移淋巴结55枚,转移率为47.06%,转移度为22.82%。结论微创手术能够取得与传统开胸手术一致的淋巴结清扫效果,在胸段食管癌的临床治疗方面具有较高可操作性。     

Prognostic role of KiSS-1 and possibility of therapeutic modality of metastin, the final peptide of the KiSS-1 gene, in urothelial carcinoma

The KiSS-1 gene has been reported to be a metastasis suppressor gene in human melanoma. The gene product was isolated from human placenta as the ligand of GPR54, a G protein-coupled receptor, and the C-terminally amidated peptide of 54 amino acids is called metastin. The binding of metastin to GPR54 has been shown to inhibit tumor metastasis in some tumor cells; however, its function remains unclear in urothelial carcinoma. We first evaluated KiSS-1 expression and GPR54 expression in 151 patients with upper urinary tract urothelial carcinoma to determine their prognostic significance. Next, we examined the role of metastin in the invasiveness and lung metastasis of MBT-2 variant (MBT-2V), which is a highly metastatic murine bladder cancer cell. Multivariate analysis revealed that KiSS-1 expression was an independent predictor of metastasis and overall survival. However, GPR54 expression was not selected. Hematogeneous metastasis had a significantly lower level of KiSS-1 expression compared with lymph node metastasis. Metastin treatment significantly reduced the invasiveness of MBT-2V cells and inhibited the DNA-binding activity of NF-κB by blocking its nuclear translocation, leading to a reduction in the expression and activity of matrix metalloproteinase-9. Metastin treatment dramatically prevented the occurrence of lung metastatic nodules (6.3 ± 2.3, n = 15) compared with controls (30.4 ± 5.1, n = 15; P < 0.01), as well as had survival benefit. KiSS-1 plays an important role in the prognosis of upper tract urothelial carcinoma and metastin may be an effective inhibitor of metastasis in urothelial carcinoma through its blockade of NF-κB function.     

乳腺浸润性导管癌c-erbB-2的表达与淋巴结转移和肿瘤细胞分裂能力的关系

目的探讨乳腺浸润性导管癌中c-erbB-2与肿瘤生物学特性的关系。方法应用免疫组化SP方法对检测乳腺浸润性导管癌c-erbB-2的表达情况极其淋巴结转移和癌细胞分裂能力的相关性。结果c-erbB-2阳性表达主要分布在肿瘤细胞的胞膜上;随着c-erbB-2表达强度的降低,肿瘤淋巴结转移率呈减少的趋势;肿瘤组织中细胞的核分裂数目随着c-erbB-2的表达强度增加而增多。结论c-erbB-2是乳腺癌细胞增殖、转移能力和预后判定的参考指标。     

进展期胃癌淋巴结转移的螺旋CT征象与病理学检查相对照

目的 探讨胃癌淋巴结转移的螺旋CT征象及其与病理学的关系.方法 对59例进展期胃癌患者行螺旋CT平扫及三期动态增强扫描. 结果①胃癌淋巴结螺旋CT呈融合型、强化明显、不均匀强化及直径≥9 mm者,病理转移的阳性率较高(P<0.05);②胃癌组织呈低分化、Borrmann Ⅲ+Ⅳ、浸润深度T3～4、TNM分期Ⅲ+Ⅳ者,螺旋CT诊断淋巴结转移的阳性率较高(P<0.05),而与肿块的大小无关. 结论 胃癌淋巴结的螺旋CT征象(淋巴结分布类型、大小、强化程度、强化方式)可反映转移淋巴结的病理特性;胃癌组织的病理学特性(癌组织的分化程度、Borrmann分型、浸润深度及TNM分期)决定螺旋CT判定淋巴结转移的阳性率.综合胃癌组织及淋巴结的各种螺旋CT征象有助于提高CT诊断淋巴结转移的准确性.     

Measurement of cytotoxic activity in experimental cancer

Several tumor cell systems have been developed for investigating the cytotoxic activity of different substances. The Ehrlich ascytic tumor (EAT) has been studied extensively and used to increase our comprehension of immunotherapy (Yamamoto and Naraparaju. Cancer Res 57:2187-2192, 1997). In this study, we evaluated the ability of an enzymatic method to assess cytotoxic tumor activity in vitro. To prepare cytotoxic cells, lymphoid cells isolated from the lymph nodes of C57BL/6 mice were activated with interleukin-2 (IL-2). The cytotoxic activity to EAT cells was assessed by an enzymatic test using lactate dehydrogenase (LDH), an enzyme widely distributed in mammalian tissues which, in the presence of NAD/NADH, converts lactate to pyruvate. IL-2 treatment of lymph node cells induced a greater percentage of lysis (75.47%) than effector cells that were not treated with IL-2 (42.38%). This suggests that IL-2 directly activates effector cells and not tumor cells. The results demonstrate that the LDH release-measurement method can be utilized to assay cytotoxic cell-mediated activity in which murine tumor cells act as the target. Levels of IL-2 appear to have a direct correlation to cellular immunity and treatment with these substances may strengthen immune function and decrease the pace of disease development.     

Contrast-aided diagnostic ultrasound does not enhance lung metastasis in a mouse melanoma tumor model.

OBJECTIVE: The purpose of this research was to test the hypothesis that contrast-aided diagnostic ultrasound (CADUS) could exacerbate the metastatic spread of mouse melanoma tumor cells to the lungs. METHODS: The melanoma cell lines B16 and B16-D5 (metastatic specifically to lung) were implanted on a hind leg of female C57/bl6 mice. Growing tumors were scanned by 1.5-MHz diagnostic ultrasound in a 37 degrees C water bath. Four hundred image frames were triggered at a 1-Hz rate with 4 retro-orbital injections of an ultrasonographic contrast agent at dosage of 10 microL/kg at 100-second intervals. Sham-treated mice received 400 frames of ultrasonography followed by the contrast agent with the ultrasound off. The primary tumor was surgically removed 1 day after ultrasound administration. Lungs were removed and evaluated blind after 2 weeks of bleaching in Fekete solution. RESULTS: Three experiments were performed. The first experiment involved scanning sham and CADUS groups of 20 mice each with B16 tumors; B16 metastasis was not enhanced. The second experiment repeated this test with the D5 cell line; the metastasis enhancement was marginally significant for average number (0.3 and 3.2; P = .06) and incidence (3 and 9 of 19; P = .08) in mice without tumor recurrence. Finally, a third experiment was performed to clarify ambiguous results in the second experiment and consisted of 2 groups of 40 mice each. In this larger experiment, the results were essentially equal for the sham and CADUS groups. CONCLUSIONS: Overall, the results do not support the hypothesis of CADUS-enhanced metastasis.     

VEGF-A induces tumor and sentinel lymph node lymphangiogenesis and promotes lymphatic metastasis

The mechanisms of tumor metastasis to the sentinel lymph nodes are poorly understood. Vascular endothelial growth factor (VEGF)-A plays a principle role in tumor progression and angiogenesis; however, its role in tumor-associated lymphangiogenesis and lymphatic metastasis has remained unclear. We created transgenic mice that overexpress VEGF-A and green fluorescent protein specifically in the skin, and subjected them to a standard chemically-induced skin carcinogenesis regimen. We found that VEGF-A not only strongly promotes multistep skin carcinogenesis, but also induces active proliferation of VEGF receptor-2-expressing tumor-associated lymphatic vessels as well as tumor metastasis to the sentinel and distant lymph nodes. The lymphangiogenic activity of VEGF-A-expressing tumor cells was maintained within metastasis-containing lymph nodes. The most surprising finding of our study was that even before metastasizing, VEGF-A-overexpressing primary tumors induced sentinel lymph node lymphangiogenesis. This suggests that primary tumors might begin preparing their future metastatic site by producing lymphangiogenic factors that mediate their efficient transport to sentinel lymph nodes. This newly identified mechanism of inducing lymph node lymphangiogenesis likely contributes to tumor metastasis, and therefore, represents a new therapeutic target for advanced cancer and/or for the prevention of metastasis.     

Enhanced tumorigenesis and lymphatic metastasis of CD133+ hepatocarcinoma ascites syngeneic cell lines mediated by JNK signaling pathway in vitro and in vivo

Cancer stem cells (CSCs), stem-like cells, or tumor-initiating cells (TICs) may initiate tumorigenesis and metastasis, but neither the basic cell biology of CSCs nor the mechanisms of CSC-mediated tumor growth and lymphoid node metastasis are understood. Evidence suggests that CSC phenotype is maintained, at least in part, by altered JNK signaling. In this study, factors influencing the growth and metastatic potential of CSCs were examined by comparing CD133 surface antigen expression, proliferation, clonogenicity, invasive capacity, tumorigenicity, and expression of JNK-associated signaling molecules between the highly metastatic mouse hepatocarcinoma ascites syngeneic cell line Hca-F and the low metastasis potential line Hca-P. The Hca-F line exhibited higher clonogenic, proliferative, and invasive capacities than Hca-P cells, and a greater proportion of Hca-F cells were CD133 positive. In both cell lines, the CD133+ subpopulation showed significantly enhanced tumorigenicity and metastatic potential. An in vivo tumorigenicity assay in nude mice indicated that Hca-F cells possessed significantly higher tumorigenicity than Hca-P cells as indicated by larger tumors after inoculation. Expression levels of E-cadherin (CDH1), annexin VII, and JNK1 proteins were inversely correlated with CD133 expression in both Hca-F and Hca-P cells. These results demonstrate that CD133+ subpopulations of both Hca-F and Hca-P lines show CSC-like properties. However, Hca-F cells showed greater tumorigenicity and invasiveness, consistent with greater lymphatic metastasis capacity. We propose that tumorigenesis and lymphatic metastasis are regulated by JNK/P53/annexin VII and JNK/ATF-2/CDH1/annexin VII signal transduction pathways.     

Human cytotoxic T lymphocytes evoked by group A streptococcal M proteins

Purified group A streptococcal M proteins were used to stimulate peripheral blood lymphocytes from normal adult volunteers. The activated lymphocytes were cytotoxic against cultured human heart cells, as well as liver cells, fibroblasts, and K562 cells, but showed only minimal cytotoxicity against several animal cell types. The cytotoxic activity evoked by type 5 M protein was dose and time dependent. Rabbit antisera against pep M5 that contained heart-crossreactive antibodies partially inhibited cytotoxicity against heart cells, but had no effect on other target cells, suggesting that a fraction of the effector lymphocytes may be recognizing M protein-crossreactive cell surface antigens. All of the cytotoxic activity was recovered from a CD3+ population of lymphocytes obtained from a fluorescence-activated cell sorter, and CD4+ and CD8+ cells were also cytotoxic. M protein-responsive T cell clones were generated that showed specificity for heart and K562 cells, in addition to clones that were cytotoxic against both cell lines. Our data show that streptococcal M protein evokes cytotoxic T lymphocytes against multiple human but not animal target cells. Some of the effector cells may be specific for cultured myocardial cells, but their role in the pathogenesis of rheumatic carditis will require further studies of lymphocytes from patients with acute rheumatic fever and carditis.     

Suppression of tumor formation in lymph nodes by L-selectin-mediated natural killer cell recruitment

Natural killer (NK) cells are known to reject certain tumors in vivo; however, the ability of NK cells to prevent metastasis of tumors into secondary lymphoid organs has not been addressed. Here, we report that in tumor-bearing hosts, NK cells are recruited to regional lymph nodes in wild-type mice, but not in mice deficient for L-selectin or L-selectin ligands. By adoptive transfer and complete Freund's adjuvant stimulation experiments, we demonstrated that L-selectin on NK cells and L-selectin ligands on endothelial cells are essential for NK cell recruitment to lymph nodes. Furthermore, freshly isolated resident lymph node NK cells lysed tumors efficiently, and metastasis of B16 melanoma cells to draining lymph nodes was suppressed in wild-type or Rag-1-deficient mice, but not when NK cells were depleted. Although L-selectin-deficient NK cells efficiently lysed tumor cells in vitro, NK cell-dependent suppression of tumor metastasis was diminished in mice deficient for L-selectin or L-selectin ligands because of insufficient NK cell recruitment to lymph nodes. Moreover, tumor metastasis was substantially inhibited in L-selectin-deficient mice reconstituted with wild-type NK cells. These findings indicate that L-selectin-mediated NK cell recruitment plays a crucial role in the control of tumor metastasis into secondary lymphoid organs.     

能量多普勒超声检测乳腺癌血管生成活性及其与腋窝淋巴结转移的关系

目的　研究乳腺癌血管生成活性与腋窝淋巴结转移的关系。方法　采用能量多普勒 (PDI)检测80例乳腺癌内血流信号并通过图像分析技术定量测定肿瘤内血管阳性反应总面积 ,同时采用免疫组化技术以FVIII RA检测肿瘤内微血管密度 (MVD) ,分析两者与腋窝淋巴结转移的关系。结果　有腋窝淋巴结转移即LN( )组血流信号较无腋窝淋巴结转移即LN(-)组明显丰富 ,至少可见 1支以上穿入型血管束 ,LN( )组血管阳性反应总面积 (40 0 9± 2 6 8)明显高于LN(-)组 (2 116± 2 0 0 ) ,差异有显著性意义 (P <0 .0 1) ,LN ( )组MVD(5 7.14± 30 .98)较LN (-)组MVD(2 3 .96± 12 .96 )亦显著增高 (P <0 .0 1)。结论　乳腺癌血管生成活性与其腋窝淋巴结转移密切相关 ,血管生成活跃者 ,腋窝淋巴结转移可能性大。PDI对乳腺癌肿瘤组织内血流的定量检测可能为临床判断乳腺癌预后提供一较为简便的指标。     

口腔鳞状细胞癌颈淋巴结转移术前评价

目的 :准确评价口腔鳞状细胞癌 (oralsquamouscellcarcinomaOSCC)颈淋巴结转移。方法 :采用临床TNM分类、组织病理分级、肿瘤生长类型、淋巴细胞浸润、肿瘤浸润厚度等对 6 4例OSCC进行观察多因素分析。结果 :TNM分类、组织病理分级、肿瘤生长类型、淋巴细胞浸润程度不能准确反     

胸中下段食管鳞癌淋巴结转移趋势的研究

目的了解胸中下段食管鳞癌淋巴结转移趋势,探讨合理的淋巴结清扫范围。方法回顾性分析浙江省肿瘤医院胸腹外科收治的933例胸中下段食管鳞癌患者的临床及病理资料,应用χ^2检验进行样本率及构成比的比较。结果933例患者中517例经病理学证实存在淋巴结转移（转移率55．4％）,其中320例单独或合并存在腹腔淋巴结转移。全组总共清除淋巴结26118枚,转移2142枚（转移率8．2％）,其中腹腔淋巴结清除12072枚,转移906枚。不同分段食管鳞癌淋巴结转移方向分布不同（χ^2=7．90,0．01〈P〈0．05）,胸中段上行与下行转移的频度相当,胸下段下行转移多见,且胸下段腹腔淋巴结转移率及转移度均高于胸中段（χ^2=52．83,P〈0．01;χ^2=134．52,P〈0．01）。除7例Tis期食管鳞癌未发生淋巴结转移外,T,以后各期均可见淋巴结转移。不同T分期胸中下段食管鳞癌淋巴结转移方式分布不同（χ^2=18．12,0．05〈P〈0．01）,病变浸润越深,发生连续性转移的机会越多,而病变浸润越浅,发生跳跃性转移的机会越多。结论胸中下段食管鳞癌淋巴结清扫范围应参照淋巴结转移趋势,结合术前检查结果,合理地选择。同时,应重视腹腔淋巴结的清扫,尤其胸下段癌。