Relationship between genetic instability of nm23H1 gene and clinical pathological characteristic in hepatocellular carcinoma

AIM: The aim of this study was to examine the microstatellite instability (MSI) and loss of heterozygosity(LOH) of locus D17S396 on chromosome 17 and their influence on the expression of nm23H1 in hepatocellular carcinoma (HCC),which may provide experimental evidence for the mechanism of nm23H1 gene and tumor metastasis.METHODS: Techniques such as DNA extraction from formalin-fixed paraffin-embedded tissues,PCR-SSCP,ordinary silver stain were used to study MSI and LOH of locus D17S396.Envision immunohistochemistry and Leica-Qwin computer imaging techniques were used to assess the expression of nm23H1.RESULTS: ① The frequency of heredity instability of HCCs was 35.42%.The frequency of LOH in the cases with lymph node or distant organs metastasis or not and with intrahepatic metastasis or embolus of portal vein or not was significantly different (P<0.01),it was higher in stage TNM Ⅲ than that in stageⅠ and Ⅱ.Moreover,it was higher in high tendency to invasion or metastasis cases than that in the low tendency cases (P<0.01).② The expression of nm23H1 was 56.25%.It was significantly different in Edmondson grade,TNM stage and in lymph node or distant organ metastasis cases (P<0.01).The cases with high tendency of invasion or metastasis exhibited lower nm23H1 expression compared with low tendency cases (P<0.01).③ The positive rate of nm23H1 protein in LOH positive group was lower than that in LOH negative group (P<0.05).CONCLUSION: The results indicate that both MSI and LOH of nm23H1 gene control the development of HCC independently in different pathways.LOH inhibits the expression of nm23H1,which endows it with high aggressive and poor prognosis.Increase in the amount of nm23H1 protein expression effectively restrains the tendency to invasion or metastasis of HCCs and improves prognosis of patients.     

Study on genetic instability of nm23-H1 gene and expression of hMLH1,hMSH2 in sporadic gallbladder carcinoma

AIM: To examine the MSI and LOH of locus D17S396 and their influence on the expression of nm23-H1 in gallbladder tumors,and to examine the protein expression of hMLH1/hMSH2,which may provide experimental evidence for the tumor occurrence and metastasis.METHODS: Techniques such as DNA extraction,CR-SSCP,ordinary silver stain were used to study MSI and LOH of locus D17S396.Envision IHC was used to assess the expression of nm23-H1 and hMLH1/hMSH2.RESULTS: ① The frequency of heredity instability of gallbladder carcinoma was 42.55%.The frequency of LOH in liver and lymph node metastasis cases and in stage Nevin IV and V was significantly higher than that without metastasis and stage I,II and III.However,the frequency of MSI showed contrary correlation with some clinicopathologic characteristics.② The expression of nm23-H1 was 46.81%.The case with lymph node metastasis and Nevin stage IV and V showed significantly lower expression than that without lymph node metastasis and stage I,II and III.③ The expressions of hMLH1 and hMSH2 were 51.06% and 42.55% respectively.hMLH1 in lymph node and liver metastasis cases and in stage Nevin IV and V were significantly lower than that without metastasis and in stage I,II and III.④ Positive frequency of hMLH1 in MSI positive group was higher than that in MSI negative group.The positive frequency of nm23-H1 and hMSH2 protein in LOH positive group was lower than that in negative group.CONCLUSION: The heredity instability of nm23-H1 gene may be implicated pathogenesis and progression of gallbladder carcinoma.Both MSI and LOH of nm23-H1 control the development of gallbladder carcinoma independently in different paths.Abnormal expression of hMLH1/hMSH2 may be a molecule marker in early stage of gallbladder carcinoma.     

Expression and prognostic significance of SLP-2 in gastric cancer

AIM: To investigate the expression of the red cell membrane integration protein SLP-2 (stomatin-like protein 2) in gastric cancer tissues and to analyze its correlation with clinicopathological manifestations and prognosis. METHODS: One hundred and ninety gastric cancer tissue samples with detailed clinical information were collected from the Department of Pathology, Sun Yat-sen University Cancer Center. The protein expression of SLP-2 in ganstric cancer was detected by the method of immunohistochemistry. The relationships between SLP-2 expression and the clinicopathological manifestations were evaluated. RESULTS: The positive rate of SLP-2 in gastric cancer tissue was 63.2% (120/190). SLP-2 expression was relevant to infiltration depth, TNM stage and lymph node metastasis (P<0.05). However, no statistical difference was observed in the SLP-2 expression associated with sex, age, differentiation, tumor size and distant metastases. Kaplan-Meier survival curves revealed that increased expression of SLP-2 was associated with poor prognosis in gastric adenocarcinoma patients (P<0.01). Based on the univariate analysis, 7 factors were found to have statistical significance of associations with overall survival, including SLP-2 expression, lymph node metastasis, histological grade, tumor size, invasive depth, distant metastases and the 7th edition of the UICC TNM classification. Only the tumor size and the 7th edition of the UICC TNM classification were independent prognostic factors for overall survival in the multivariate analysis. CONCLUSION: SLP-2 is highly expressed in gastric adenocarcinoma tissues and may play an important role in tumor progression and metastasis. Although SLP-2 is not an independent prognostic factor, it may influence the prognosis of gastric cancer. Increased expression of SLP-2 can be used for predicting unfavorable prognosis in gastric adenocarcinoma patients.     

The microsatellite instability and loss of heterozygosity in head and neck squamous cell carcinomas

AIM:To examine the microsatellite instability (MSI) and loss of heterozygosity (LOH) in head and neck squamous cell carcinomas (HNSCC). METHODS: 36 cases of HNSCC were analyzed with 15 microsatellite markers from chromosome 3,5,6,8,9,13,17 and 18. RESULTS: Among the 36 cases of HNSCC, 27.8%(10/36)of samples showed MSI in one to eight microsatellite markers. High frequent MSI occurred at D17S520(22.9%), D6S105(16.7%)and D8S264(13.9%). LOH was detected on the site of 9p21-p22 and 3p14. No correlations were found between allelic instability and grade or stage of the tumor. CONCLUSION: Our data suggest that MSI is a common genetic change in HNSCC. Tumor suppressor genes related to HNSCC may harbor at chromosome 9p21-p22 and 3p14 regions.     

Study of annexin A2 expression in gastric cancer by proteomics and tissue microarray

AIM: To investigate the differential expression of annexin A2 (ANXA2) in gastric carcinoma and to analyze the relationship between ANXA2 expression and clinicopathological parameters of gastric carcinoma. METHODS: Pure gastric adenocarcinoma cells (GAC) and normal gastric epithelial cells (NGEC) in 15 patients with gastric cancer were acquired by laser capture microdissection (LCM). All peptide specimens after trypsin digestion were labeled with ¹⁸O/¹⁶O. Quantitatively identification of differential expression of the proteins betweem GAC and NGEC was performed by Nano-RPLC-MS/MS. The expression of ANXA2 in the 2 kinds of tissues was detected by Western blotting. Tissue microarray containing 75 pairs of gastric carcinoma and para-carcinoma tissues was used and the expression of ANXA2 in these specimens was detected by the method of immunohistochemistry (IHC). The relationship between ANXA2 expression and clinicopathological parameters of the pateints with gastric carcinoma was analyzed. RESULTS: A total of 78 differential proteins were identified and ANXA2 was up-expressed in GAC (2.32∶ 1), which was confirmed by Western blotting (P<0.01). The results of IHC showed that the correlations between the expression level of ANXA2 protein and invasive depth (T stage), lymph node metastasis (N stage), histological differentiation, TNM stage and the size of tumor were observed (P<0.01), but the correlations between the ANXA2 expression and sex, age and distant metastasis (M stage) were not found (P>0.05). CONCLUSION: The up-expressed ANXA2 may play an important role in the biological behavior of gastric cancer.     

Expression of CD44s in lung cancer and its significance

AIM: To investigate expression of CD44s in lung cancer and it's clinical significance. METHODS: A total of 117 primary lung cancer from patients were examined for CD44s expression by immunohistochemical staining. RESULTS: CD44s mostly expressed in non-small cell lung cancer (NSCLC) but not in small ecll lung cancer (SCLC), and squamous cell carcinoma(SCC) showed much stronger expression of CD44s than adenocarcinoma(ADC)(P＜0.05). In comparison of the lung cancer with/ without lymph node metastasis, the latter showed stronger expression of CD44s(P＜0.01). According to TNM, there was a distinct statistic difference between early stage and advanced stage(P＜0.05). CONCLUSION: CD44s might be a better indicant in histological classification of lung cancer, lymph node metastasis, clinical stage and prognosis.     

Correlation between E-cadherin and FOXO3a expression in gastric can-cer tissues and cells

AIM: To investigate the expression of E-cadherin and forkhead box protein O3a (FOXO3a) in gastric cancer tissues and cells, and its correlation with cell viability. METHODS: The expression of E-cadherin and FOXO3a was detected by immunohistochemical staining in 53 specimens of gastric cancer tissues and their adjacent tissues, and the relationship between their expression and clinicopathological characteristics were analyzed. E-cadherin-over-expressing gastric cancer AGS cells were constructed by lentivirus-mediated cell transfection, and the protein expression of E-cadherin and FOXO3a was detected by immunocytochemistry method. The expression of E-cadherin, FOXO3a, Akt, Bcl-2 and Bax was determined by Western blot. The cell viability was detected by CCK-8 assay. RESULTS: The positive expression rates of E-cadherin and FOXO3a proteins in gastric cancer tissues were both significantly lower than those in their adjacent tissues (P<0.05). E-cadherin positive expression in gastric cancer tissues was significantly related to tumor grade and TNM stage (P<0.05), but not related to age, sex, location, T stage or lymph node metastasis. FOXO3a positive expression was significantly related to tumor grade (P<0.05), but not related to age, sex, location, TNM stage, T stage or lymph node metastasis. The expression of E-cadherin was positively correlated with FOXO3a expression in gastric cancer tissues (r=0.376, P=0.003). After over-expression of E-cadherin, the viability of gastric cancer AGS cells was significantly inhibited, the expression of FOXO3a, Bcl-2 and Bax was significantly increased, and the expression of Akt was significantly decreased. CONCLUSION: E-cadherin and FOXO3a are involved in the development of gastric cancer, and E-cadherin may affect the viability of gastric cancer cells by regulating Akt/FOXO3a signaling pathway.     

Significance of STC1 expression in peripheral blood and tissue of gastric carcinoma

AIM: To investigate stanniocalcin 1 (STC1) mRNA expression in the blood and tissue of gastric carcinoma. METHODS: The samples of peripheral blood and tumor tissues were collected from 74 patients with stage I-IV gastric carcinoma. STC1 mRNA was detected by RT-PCR, STC1 protein in tumor tissue was also detected by immunohistochemical method. RESULTS: The expression of STC1 mRNA in blood was significantly correlated with multiple histopathological prognostic factors, including primary tumor size, number of positive lymph nodes, and TNM stage. STC1 mRNA was not detected in the blood of volunteers without cancer, but detected in all gastric carcinoma tissue, and STC1 protein was expressed in all gastric carcinomas. CONCLUSION: STC1 is proposed as a novel, specific, and clinically useful molecular marker for detecting occult gastric carcinoma cells in blood. STC1 expression may play an important role in the progression of gastric carcinoma.     

Expression and clinical significance of Bmi-1 in gastric carcinoma

AIM: To investigate the relationship between expression of Bmi-1 (B cell-specific MLV integration site-1) in gastric cancer and its clinicopathologic significance.METHODS: 146 surgical patients with gastric carcinoma were followed up at least 2 years.Expression of Bmi-1 protein was examined by immunohistochemistry in their archival paraffin embedded tissue specimens.RESULTS: The intensive positive rate of Bmi-1 expression in gastric cancer was 67.8% (99/146).Expression of Bmi-1 was highly correlated with tumor size,clinical stage,lymph node metastasis and T classification (P<0.05),but not with sex,age,tumor differentiation,etc.(P>0.05).The survival rate in the patients with Bmi-1 expression was much lower than that in those patients without Bmi-1 expression (P<0.01).Multivariate analysis indicated that Bmi-1 expression,T classification,lymph node metastasis,distant metastasis,tumor size and postoperative chemotherapy were all significantly prognostic factors of gastric carcinoma.CONCLUSION: Overexpression of Bmi-1 in patients with gastric carcinoma enhances the possibility of invasion and metastasis,implying a poor prognosis.Bmi-1 may serve as fairly a good prognostic factor to indicate biologic behavior and prognosis in gastric carcinoma.     

Expression of bFGF in malignant tumor and its clinical pathological significance

AIM: To detect basic fibroblast growth factor (bFGF) expression in clinical common malignant tumor (non-small-cell lung cancer,breast cancer, colon cancer and melanoma), and to identify relationship between the expression and tumor clinicopathological characteristics.METHODS: Immunohistochemical SP method was used to detect the expression of bFGF at protein level in 208 cases of paraffin-embedded tissue of primary malignant tumor patients (68 cases of lung cancer, 80 cases of breast carcinoma, 41 cases of colon cancer and 19 cases of melanoma).RESULTS: The bFGF protein expression levels were significantly higher in low differentiated non-small-cell lung cancer with lymph node metastasis, and were positively correlated with TNM. In addition, no significant influence of the bFGF protein expression on the patients with median survival period was observed. The protein expression of bFGF was higher in advanced breast cancer with lymph node metastasis and was commonly found in the middle/higher differentiated colon cancer with regional lymph node metastasis. Meanwhile, bFGF protein was highly expressed in advanced melanoma patients with lymph node metastasis.CONCLUSION: bFGF may participate in the process of occurrence and progression of malignant tumor. Expression of bFGF protein may be an effective parameter for evaluating metastasis and prognosis of malignant tumor.     

Expression and clinical significance of RhoC and Ki-67 in human esophageal squamous cell carcinoma tissues

AIM: To investigate the expression and significance of RhoC and Ki-67 in human esophageal squamous cell carcinoma (ESCC) tissues.METHODS: The expression of RhoC and Ki-67 was detected in 52 specimens of ESCC by the method of immunohistochemistry. The clinicopathological features were also analyzed.RESULTS: The expression of RhoC was detected in 32 of the total 52 (61.5%) cases of human ESCC tissues, significantly higher than that in the adjacent histologically normal epithelium, which was only in 11 of 37 cases (29.7%, P<0.05). RhoC expression was closely correlated with clinical tumor-node-metastasis (TNM) stage (P<0.05) and lymph node metastasis (P<0.05) in ESCC. The over-expression of RhoC was positively correlated with Ki-67 in ESCC (r=0.322, P<0.05).CONCLUSION: The over-expression of RhoC protein significantly correlates with advanced TNM stage, lymph node metastasis and cell proliferation ability of ESCC. Therefore, RhoC may be a new auxiliary parameter for early diagnosis and prognostic evaluation of ESCC.     

Central role of GRP78 in growth of gastric carcinoma cells

AIM: To explore the effect of glucose-regulated protein 78 (GRP78) on the gastric carcinogenesis. METHODS: GPR78 expression patterns were examined in 34 specimens from gastric carcinoma patients using the immunohistochemistry (IHC) assay, and in 10 specimens using Western blotting analysis. In addition, the expression of GPR78 and cyclin D1 was detected in human gastric cancer cell lines SGC7901 and SGC7901-H78 (overexpressing GRP78) by Western blotting. RESULTS: By IHC assay, GRP78 was found to be highly expressed in the cytoplasm of gastric carcinomas as compared with the adjacent non-malignant tissues and corresponding normal tissues. GRP78 expression was positively correlated with gender and histological differentiation (P<0.05), but not with age, tumor stage and lymph node metastasis (P>0.05). Furthermore, we found that with the increased expression of GRP78 in SGC7901-H78 cells, the expression of cyclin D1 was also elevated. CONCLUSION: GRP78 might be a key player to be involved in the growth of gastric cancer.     

Methylation and mRNA expression of TIG1 gene in esophageal squamous-cell carcinoma tissues

AIM: To investigate the expression and promoter methylation of tazarotene-induced gene-1 (TIG1) in esophageal squamous-cell carcinoma (ESCC) tissues. METHODS: The methods of methylation-specific PCR and real-time fluorescence quantitative PCR were applied to examine the methylation and mRNA expression of TIG1, respectively, in 43 cases of ESCC tissues, 20 cases of paracancerous tissues and 15 cases of normal tissues. RESULTS: The frequency of promoter methylation of TIG1 gene in ESCC tissues was 25.6% (11/43), which was significantly higher than that in the paracancerous tissues (5.0%, 1/20) and normal tissues (0/20). The hypermethylation of TIG1 gene in these tissues had no correlation with sex, age and clinical stage of the patients. However, it was correlated with the pathological stage (P<0.01) and lymph node metastasis (P<0.05). The mRNA expression of TIG1 in ESCC tissues was significantly lower than that in paracancerous tissues (P<0.05) and normal tissues (P<0.01). However, the expression level of TIG1 mRNA in methylated tissues was significantly lower than that in unmethylated tissues (P<0.01). CONCLUSION: Promoter methylation may be an important mechanism of TIG1 gene inactivation in ESCC, which was related to lymph node metastasis and TNM stage of esophageal carcinoma.     

Expression of miR-143 and its clinicopathologic significance in gastric cancer

AIM: To investigate the expression of miR-143 and its association with clinicopathologic features in gastric cancer.METHODS: The expression level of miR-143 in 32 cases of gastric cancer and matched non-tumor adjacent tissue specimens was examined using stem-loop real-time RT-PCR. The relationship between the expression of miR-143 and its clinicopathologic features of gastric cancer was analyzed.RESULTS: The expression level of miR-143 was significantly lower in the tumor tissues than that in the adjacent tissues (P<0.05). Down-regulated miR-143 expression was associated with the cell differentiation (P<0.05) and lymph node metastasis (P<0.05) in gastric cancer patients. No significant association was found between the expression of miR-143 and the status of gender, age, blood type, tumor location, tumor size, depth of tumor invasion and tumor node metastasis stage.CONCLUSION: It is possible that miR-143 plays an important role in the generation and progression of gastric cancer. The expression level of miR-143 may be a valuable adjuvant parameter for predicting the poorly differentiated gastric cancer.     

High expression of BIRC5 enhances cell viability,inhibits apoptosis and is associated with poor prognosis in gastric cancer

AIM To investigate the expression of baculoviral inhibitor of apoptosis protein repeat-containing protein 5 （BIRC5） in gastric cancer tissue and its relationship with prognosis of gastric cancer patients, and to explore the effect of BIRC5 knock-down on the viability and apoptosis of gastric cancer cells. METHODS The expression of BIRC5 was detected by immunohistochemistry in 67 cases of gastric cancer tissues and paracancerous tissues for analyzing the relationships with clinicopathological characteristics. The mRNA and protein expression levels of BIRC5 in gastric carcinoma cell lines （AGS, MKN-1 and MGC-803） and normal gastric epithelial cell line GES-1 were detected by RT-qPCR and Western blot. The AGS cells were divided into blank group （no treatment）, Ctr-sh group （blank plasmid transfection） and BIRC5-sh group （BIRC5-shRNA plasmid transfection）. The interference efficiency of BIRC5-shRNA was evaluated by Western blot. The cell viability was measured by MTT assay, the apoptosis was analyzed by flow cytometry, and the levels of apoptosis-related proteins cleaved caspase-3, Bax and Bcl-2 were determined by Western blot. RESULTS BIRC5 was mainly expressed in cytoplasm, and the positive expression rate of BIRC5 in the gastric cancer tissues was higher than that in the adjacent tissues （P<0.01）. The positive rates of BIRC5 in the gastric cancer patients at TNM Ⅲ~Ⅳ stages and with lymph node metastasis were higher than those in the patients at TNM Ⅰ~Ⅱ stages and without lymph node metastasis, respectively （P<0.05）. The survival time of the patients with positive BIRC5 expression was shorter than that of the patients with negative BIRC5 expression （P=0.011 2）. The cell viability in BIRC5-sh group was lower than that in blank group and Ctr-sh group at time points of 48, 72 and 96 h. The apoptotic rate in BIRC5-sh group was increased compared with blank group and Ctr-sh group. The protein levels of cleaved caspase-3 and Bax in BIRC5-sh group were higher than those in blank group and Ctr-sh group, while the protein expression of Bcl-2 in BIRC5-sh group was lower than that in blank group and Ctr-sh group （P<0.05）. CONCLUSION High expression of BIRC5 in gastric cancer indicates poor prognosis. BIRC5 promotes the growth of gastric cancer cells and inhibits apoptosis.     

Relationship between c-erbB-2, BCSG1 expression and recurrence,metastasis in breast cancer

AIM: To assess the significance of c-erbB-2, BCSG1 (breast cancer specific gene-1) expression and other parameters in recurrence or metastasis of breast cancer. METHODS: The expression of c-erbB-2, BCSG1, and ER, PR, MVD, VEGF, VEGF-C, FLT-4, LVD were determined with the SP immunohistochemical method in 58 cases of invasive breast cancer patients occurred over 5 years. The cases were used to analyze the effect of c-erbB-2, BCSG1, VEGF-C and ER, PR, MVD, VEGF, FLT-4, LVD expression on clinical-pathological manifestations and prognosis in breast cancer. RESULTS: The expression rates of c-erbB-2, BCSG1, VEGF-C, LVD were respectively 25.9%, 62.1%, 36.2%, 32.8% in association with the lymph node metastasis and recurrence of breast cancer (P<0.05), the expression rate of MVD was also increased significantly (P＜0.05). CONCLUSION: The c-erbB-2, BCSG1, VEGF-C, LVD are highly expressed and strongly correlated with the lymph node metastasis and recurrence of breast cancer, of which BCSG1 may be used as a predictor of prognosis.     

Relationship between RUNX3,cyclin E,P21 and survival in gastric cancer patients

AIM:To evaluate the relationship between RUNX3,cyclin E,P21,biological features and survival in gastric cancer patients.METHODS:RUNX3 was examined using immunohistochemical staining.Cyclin E and P21 were analyzed by flow cytometry.Survival was evaluated by Kaplan-Meier survival curves.RESULTS:The positive-expression rate of RUNX3,cyclin E and P21 in tumor tissue from 56 patients with gastric cancer were 44.6%,64.3% and 32.1%,respectively.RUNX3 expression was correlated with lymph node metastasis and distant metastasis (P<0.05).Cyclin E might be related to depth of invasion,lymph node metastasis and distant metastasis (P<0.05).P21 was correlated with lymph node metastasis (P<0.05).It was revealed that RUNX3 and P21 were correlated (r=0.57,P<0.05),no correlation between RUNX3 and cyclin E was observed (r=0.25,P>0.05).Using Kaplan-Meier survival curves and the Log-rank test,there was correlation between RUNX3,cyclin E and survival (P<0.05).No correlation between P21 and survival was observed (P>0.05).CONCLUSION:RUNX3 may be related with tumorigenesis and tumor progression by affecting P21 expression.The detection of RUNX3 and cyclin E may be helpful in evaluating the clinicopathological parameters and prognosis in gastric carcinoma patients.     

NET-1 promotes metastasis of lung squamous-cell carcinoma by activating EMT

AIM: To explore the effect of neuroepithelial cell transforming gene-1 (NET-1) expression on the metastasis of lung squamous-cell carcinoma (LSC) and the underlying molecular mechanism. METHODS: Immunohistochemistry was used to detect the expression of NET-1 protein in 53 cases of lung squamous-cell carcinoma (LSC group), 24 cases of normal lung epithelium (NLE group) and 27 cases of lung squamous intraepithelial lesions (SIL group). The correlation of clinical and pathological factors was analyzed. The protein expression of NET-1 in human lung squamous-cell carcinoma cell lines H226, H1703, H2170, SK-MES-1, H520 and YTMLC-90 was determined by Western blot. The RNA interference recombinant adenovirus against NET-1 gene (Ad-NET-siRNA) and Ad-control with control sequence were constructed and infected with human lung squamous cell carcinoma cell YTMLC-90 to silence the expression of NET-1 gene. The protein expression of NET-1, E-cadherin, vimentin and Snail1 in the BEAS-2B cells and the YTMLC-90 cells was determined by Western blot. The mRNA expression of E-cadherin and vimentin in each group of the cells was detected by qPCR. The invasive ability of the cells in each group was detected by Transwell chamber assay. RESULTS: The positive expression rate of NET-1 in LSC group was significantly higher than that in NLE group and SIL group(P<0.05). The distribution of NET-1 protein positive expression population was correlated with histological grade, lymph node metastasis, and TNM stage. The NET-1 expression rate of LSC with lymph node metastasis was significantly higher than that without lymph node metastasis. Over-expression of NET-1 protein in YTMLC-90 cells was observed. The expression of E-cadherin was decreased, and the protein expression of vimentin and Snail1 was increased in YTMLC-90 cells. Knock-down of NET-1 expression increased the expression of E-cadherin, and decreased the expression of vimentin and Snail1 in the YTMLC-90 cells. CONCLUSION: The expression of NET-1 promotes the lymphatic metastasis of lung squamous-cell carcinoma. This promotion may be achieved through the activation of epithelial-mesenchymal transition (EMT) by NET-1 expression.