LAPTM4B-35 promotes cancer cell migration via stimulating integrin beta1 recycling and focal adhesion dynamics.

Metastasis is the main cause of cancer patients' death despite tremendous efforts invested in developing the related molecular mechanisms. During cancer cell migration, cells undergo dynamic regulation of filopodia, focal adhesion, and endosome trafficking. Cdc42 is imperative for maintaining cell morphology and filopodia, regulating cell movement. Integrin beta1 activates on the endosome, the majority of which distributes itself on the plasma membrane, indicating that endocytic trafficking is essential for this activity. In cancers, high expression of lysosome-associated protein transmembrane 4B (LAPTM4B) is associated with poor prognosis. LAPTM4B-35 has been reported as displaying plasma membrane distribution and being associated with cancer cell migration. However, the detailed mechanism of its isoform-specific distribution and whether it relates to cell migration remain unknown. Here, we first report and quantify the filopodia localization of LAPTM4B-35: mechanically, that specific interaction with Cdc42 promoted its localization to the filopodia. Furthermore, our data show that LAPTM4B-35 stabilized filopodia and regulated integrin beta1 recycling via interaction and cotrafficking on the endosome. In our zebrafish xenograft model, LAPTM4B-35 stimulated the formation and dynamics of focal adhesion, further promoting cancer cell dissemination, whereas in skin cancer patients, LAPTM4B level correlated with poor prognosis. In short, this study establishes an insight into the mechanism of LAPTM4B-35 filopodia distribution, as well as into its biological effects and its clinical significance, providing a novel target for cancer therapeutics development.

[Serum LAPTM4B-35 protein as a novel diagnostic marker for hepatocellular carcinoma].

OBJECTIVE: LAPTM4B-35 protein is one of the isoforms that are encoded by a cancer driver gene, LAPTM4B. This gene was primarily found and identified in our lab of Peking University School of Basic Medical Sciences. The LAPTM4B-35 protein and its encoded mRNA are significantly over-expressed in a variety of cancers, such as hepatocellular carcinoma (HCC), lung cancers (including non small-cell lung cancer and small-cell lung cancer), stomach cancer, colorectal carcinoma, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, breast cancer, prostate cancer, ovarian cancer, cervical cancer, endometrial cancer, and so on. It has firmly demonstrated through lab experiments either in vivo or in vitro, as well as clinical studies that the over-expression of LAPTM4B-35 can promote cancer growth, metastasis, and multidrug resistance. Specially, the expressive level of LAPTM4B-35 is associa-ted with recurrence of HCC. The aim of this study is to identify the release of LAPTM4B-35 protein from hepatocellular carcinoma into blood of HCC patients and into the medium of cultured HCC cells, and to identify its possible form of LAPTM4B-35 protein existed in blood and cell culture medium, as well as to explore the possibility of LAPTM4B-35 protein as a novel HCC biomarker for diagnosis of HCC and prognosis of HCC patients. METHODS: Immunobloting (Western blot) and enzyme-linked immunosorbent assay (ELISA) were used for identification of LAPTM4B-35 protein in the blood of HCC patients and normal individuals. Ultrafiltration and ultracentrifugation were used to isolate and purify exosomes from the culture medium of HCC cells. RESULTS: LAPTM4B-35 protein existed in the blood from HCC patients and normal donors that were demonstrated through Western blot and ELISA. LAPTM4B-35 was also released into the culture medium of HCC cells in the form of exosomes. Preliminary experiments showed that the average and the median of LAPTM4B-35 protein level in the blood of HCC patients (n=43) were both significantly higher than that in the blood of normal donors (n=33) through sandwich ELISA. CONCLUSION: It is promising that the LAPTM4B-35 protein which is released from HCC cells in the form of exosomes into their extraenvironment may be exploited as a novel cancer biomarker for HCC serological diagnosis.

Serum LAPTM4B-35 protein as a novel diagnostic marker for hepatocellular carcinoma / 北京大学学报(医学版)

OBJECTIVE@#LAPTM4B-35 protein is one of the isoforms that are encoded by a cancer driver gene, LAPTM4B. This gene was primarily found and identified in our lab of Peking University School of Basic Medical Sciences. The LAPTM4B-35 protein and its encoded mRNA are significantly over-expressed in a variety of cancers, such as hepatocellular carcinoma (HCC), lung cancers (including non small-cell lung cancer and small-cell lung cancer), stomach cancer, colorectal carcinoma, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, breast cancer, prostate cancer, ovarian cancer, cervical cancer, endometrial cancer, and so on. It has firmly demonstrated through lab experiments either in vivo or in vitro, as well as clinical studies that the over-expression of LAPTM4B-35 can promote cancer growth, metastasis, and multidrug resistance. Specially, the expressive level of LAPTM4B-35 is associa-ted with recurrence of HCC. The aim of this study is to identify the release of LAPTM4B-35 protein from hepatocellular carcinoma into blood of HCC patients and into the medium of cultured HCC cells, and to identify its possible form of LAPTM4B-35 protein existed in blood and cell culture medium, as well as to explore the possibility of LAPTM4B-35 protein as a novel HCC biomarker for diagnosis of HCC and prognosis of HCC patients.@*METHODS@#Immunobloting (Western blot) and enzyme-linked immunosorbent assay (ELISA) were used for identification of LAPTM4B-35 protein in the blood of HCC patients and normal individuals. Ultrafiltration and ultracentrifugation were used to isolate and purify exosomes from the culture medium of HCC cells.@*RESULTS@#LAPTM4B-35 protein existed in the blood from HCC patients and normal donors that were demonstrated through Western blot and ELISA. LAPTM4B-35 was also released into the culture medium of HCC cells in the form of exosomes. Preliminary experiments showed that the average and the median of LAPTM4B-35 protein level in the blood of HCC patients (n=43) were both significantly higher than that in the blood of normal donors (n=33) through sandwich ELISA.@*CONCLUSION@#It is promising that the LAPTM4B-35 protein which is released from HCC cells in the form of exosomes into their extraenvironment may be exploited as a novel cancer biomarker for HCC serological diagnosis.

LAPTM4B-35 expression is associated with pathological grades and clinical stages in salivary adenoid cystic carcinoma.

Salivary adenoid cystic carcinoma is one of the most common malignancies of the head and neck. The lysosome-associated protein transmembrane-4ß gene (LAPTM4B) is a novel oncogene that has been found overexpressed in a number of clinically aggressive cancers. This study aimed to investigate the expression of the LAPTM4B-35 protein in normal salivary gland and salivary adenoid cystic carcinoma, a relatively indolent malignancy, and explore its clinicopathological significance in this malignancy. By immunohistochemical analysis, LAPTM4B-35 expression was evaluated in 106 cancer tissues, their adjacent non-cancerous tissues and five normal salivary glands. The correlation of LAPTM4B-35 expression with clinicopathological parameters was assessed using Chi-square or Fisher's exact test. The level of LAPTM4B-35 expression varied among different cell types of normal salivary glands. It was expressed at a fairly low level in serous and mucous acini, at low level in intercalated duct and excretory duct cells and moderately in secretory/striated ducts. In 50% of high grade tumor tissues tested, LAPTM4B-35 was markedly overexpressed. LAPTM4B-35 levels were significantly associated with histological grade and clinical stage. LAPTM4B-35 plays an important role in salivary adenoid cystic carcinoma and may serve as a diagnostic marker and a target for individualized therapy.

Clinical impact of circulating LAPTM4B-35 in pancreatic ductal adenocarcinoma.

PURPOSE: LAPTM4B is upregulated in a wide range of cancers associated with poor prognosis. However, the clinical impact of LAPTM4B as diagnostic and prognostic marker in pancreatic ductal adenocarcinoma (PDAC) remains unknown. Thus, the aim of the present study was to investigate the expression of LAPTM4B as circulating marker in PDAC. METHODS: Expression analysis of LAPTM4B-35 in pancreatic tissue and preoperative blood serum samples of 169 patients with PDAC UICC Stages I-IV (n = 98), chronic pancreatitis (n = 41), and healthy controls (n = 30) by immunohistochemistry, Western blot, and ELISA. Descriptive and explorative statistical analyses of LAPTM4B-35's potential as diagnostic and prognostic marker in PDAC. RESULTS: Expression of LAPTM4B-35 was significantly increased in tumor tissue and corresponding blood serum samples of patients with PDAC (each p < 0.001) and it could well discriminate PDAC from healthy controls and chronic pancreatitis (p < 0.001; p = 0.0037). LAPTM4B-35 in combination with CA.19-9 outperforms the diagnostic accuracy with an AUC of 0.903 (p < 0.001), sensitivity of 82%, and specificity of 92%. Kaplan-Meier survival analysis revealed an improved overall survival in PDAC UICC I-IV with low expression of circulating LAPTM4B-35 (17 versus 10 months, p = 0.039) as well as an improved relapse-free survival in curatively treated PDAC UICC I-III (16 versus 10 months; p = 0.037). Multivariate overall and recurrence-free survival analyses identified LAPTM4B-35 as favorable prognostic factor in PDAC patients (HR 2.73, p = 0.021; HR 3.29, p = 0.003). CONCLUSION: LAPTM4B-35 is significantly deregulated in PDAC with high diagnostic and prognostic impact as circulating tumor marker.

Prognostic Values of LAPTM4B-35 in Human Cancer: A Meta-analysis.

Background: Lysosome-associated protein transmembrane-4ß-35(LAPTM4B-35) has been observed overexpressed in multiple malignant tumors. However, the prognostic value of LAPTM4B-35 remains controversial. Therefore, we conducted a meta-analysis to evaluate the prognostic value of LAPTM4B-35 in human cancers. Methods: The relevant publications were obtained by systematically searching the PubMed, Web of Science, Embase, Wanfang, and China National Knowledge Infrastructure (CNKI) databases. Pooled hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated for the prognosis value of LAPTM4B-35 for cancer patient. Results: Our result suggest that LAPTM4B-35 overexpression is significantly associated with poor overall survival (OS) (HR = 2.49, 95% CI = 1.87-3.32, p < 0.001), disease-free survival (DFS) (HR = 2.43, 95% CI = 1.35-4.35, p = 0.003), and progression-free survival (PFS) (HR = 4.12, 95% CI = 2.30-7.37, p < 0.001). Moreover, subgroup analysis revealed significant association with poor OS in lung (HR = 2.05, 95% CI = 1.37-3.06, p < 0.001), gastric carcinoma (HR = 1.88, 95% CI = 1.01-3.50, p < 0.047) and ovarian cancer (HR = 4.94, 95% CI = 1.44-16.94, p = 0.011). Conclusion: LAPTM4B-35 may be a novel predictive biomarker and a potential target for treatment.

LAPTM4B-35 is a novel prognostic factor for glioblastoma.

Lysosome-associated protein transmembrane-4 beta (LAPTM4B)-35, a newly identified cancer-associated gene, is overexpressed in a wide variety of malignant tumors. However, studies of its expression and role in glioma have not yet been reported. This study aimed to investigate the expression and the role of LAPTM4B-35 in glioma and to assess its value as a prognostic factor. Seventy-seven glioma cases (Grade II in 18 patients, Grade III in 16 and Grade IV in 43) were immunohistochemically examined for LAPTM4B-35, pAkt, factor VIII and Ki-67 expressions. The LAPTM4B-35 expression score of Grade II gliomas was lower than those of Grade III-IV gliomas (p < 0.05), while the difference between Grade III and IV gliomas was not statistically significant. Of the 43 patients with glioblastoma (GBM), 27 (62.8%) had high LAPTM4B-35 expression, which was associated with high tumor micro-vessel density and pAkt activation. The median progression-free survival (PFS) of GBM patients with high LAPTM4B-35 expression was 5.13 months, significantly shorter than that of those with low LAPTM4B-35 expression (12.0 months, p < 0.0001). The median overall survival (OS) of GBM patients with high LAPTM4B-35 expression was 12.5 months, again significantly shorter than that of those with low LAPTM4B-35 expression (29.6 months, p < 0.0001). Multivariate analysis indicated LAPTM4B-35 to be an independent prognostic factor for PFS and OS of GBM patients. Our findings show LAPTM4B-35 to be strongly associated with tumor proliferation, tumor angiogenesis and poor outcomes of GBM patients, suggesting LAPTM4B-35 to potentially be applicable as a novel prognostic marker and even to possibly play a role in improving GBM treatment.

Expression and significance of LAPTM4B-35 and MMP-9 in patients with gastric cancer / 中国肿瘤临床

Objective:To investigate the protein expression level of lysosome-associated transmembrane protein 4β-35 (LAPTM4B-35) and matrix metalloproteinase-9 (MMP-9) in gastric cancer (GC) and to determine the relationship of LAPTM 4B-35 or MMP-9 expres-sion with clinicopathological characteristics and prognosis of patients with GC. Methods:The messenger RNA (mRNA) expression level of LAPTM4B-35 and MMP-9 was detected by real-time quantitative polymerase chain reaction, while the protein expression level of LAPTM 4B-35 and MMP-9 in GC tissues was analyzed by immunohistochemistry in formalin-fixed samples from 152 patients with GC, and data analysis was performed using statistical methods. Results:The mRNA expression level of both LAPTM4B-35 and MMP-9 in HGC-27, SGC 7901, and MGC803 GC cell lines was higher than that in the GES-1 normal gastric mucosa cell line. LAPTM 4B-35 and MMP-9 proteins mainly localized in the cytoplasm of GC cells. The positive LAPTM4B-35 expression ratio in GC tissues was 50.7％, while the MMP-9 positive ratio was 36.8％. LAPTM4B-35 and MMP-9 were positively related to gastric cancer. Lauren's classification and T stage were identified as the relative factors of LAPTM4B-35 expression, while tumor size and N stage were identified as the rela-tive factors of MMP-9 expression in GC tissues. Cox analysis indicated that the degree of differentiation, N stage, LAPTM 4B-35 expres-sion, and MMP-9 expression were independent prognostic indicators for patients with GC. Conclusion:LAPTM4B-35 and MMP-9 play important roles in the aggressiveness of GC, and LAPTM4B-35 and MMP-9 may be promising indicators for the prognosis of patients with GC.

Overexpressed LAPTM4B-35 is a risk factor for cancer recurrence and poor prognosis in non-small-cell lung cancer.

BACKGROUND: The expression levels and clinical significances of Lysosomal-associated protein transmembrane-4ß-35 (LAPTM4B-35) protein are unknown in the non-small-cell lung cancer (NSCLC). This study aimed to explore the expression and prognostic value of LAPTM4B-35 in NSCLC patients. METHODS: The clinicopathological and survival data of 107 NSCLC patients who received radical surgery from 2007 and 2011 were reviewed. The LAPTM4B-35 expression of the paired tumors and adjacent normal specimens were detected, and the association between LAPTM4B-35 and clinical variables was explored. Kaplan-Meier analysis and Cox regression (Proportional hazard model) were performed to investigate the prognostic significance for NSCLC. RESULTS: LAPTM4B-35 was over expressed in NSCLC tissues. The elevated LAPTM4B-35 expression was associated with cancer recurrence (P = 0.031). The 5-year median OS and PFS were significantly worse in the LAPTM4B-35 overexpressed group. Multivariate Cox analysis showed that LAPTM4B-35 over-expression was an independent factor for OS and PFS in NSCLC(P = 0.018, P = 0.026, respectively). CONCLUSIONS: The overexpressed LAPTM4B-35 was an independent prognostic biomarker for NSCLC, which could predict cancer recurrence and poor over survival. And that may be applied as potential target for NSCLC treatment.

Targeting a novel cancer-driving protein (LAPTM4B-35) by a small molecule (ETS) to inhibit cancer growth and metastasis.

Our previous studies demonstrated that LAPTM4B-35 is overexpressed in a variety of solid cancers including hepatocellular carcinoma (HCC), and is an independent factor for prognosis. LAPTM4B-35 overexpression causes carcinogenesis and enhances cancer growth, metastasis and multidrug resistance, and thus may be a candidate for therapeutic targeting. The present study shows ethylglyoxal bisthiosemicarbazon (ETS) has effective anticancer activity through LAPTM4B-35 targeting. Bel-7402 and HepG2 cell lines from human HCC were used as cell models in which LAPTM4B-35 is highly expressed, and a human fetal liver cell line was used as a control. The results showed ETS has a specific and pronounced lethal effect on HCC cells, but not on fetal liver cells in culture. ETS also attenuated growth and metastasis of human HCC xenograft in nude mice, and extended the life span of mice with HCC. ETS induced HCC cell apoptosis, and upregulated a large number of proapoptotic genes and downregulated antiapoptotic genes. When endogenous overexpression of LAPTM4B-35 was knocked down with RNAi, the killing effect of ETS on HepG2 cells was significantly attenuated. ETS also inhibited phosphorylation of LAPTM4B-35 Tyr285, which involves in activation of the PI3K/Akt signaling pathway induced by LAPTM4B-35 overexpression. In addition, the induction of alterations in quantity of c-Myc, Bcl-2, Bax, cyclinD1 and Akt-p molecules in HepG2 cells by LAPTM4B-35 overexpression could be reversed by ETS. CONCLUSION: ETS is a promising candidate for treatment of HCC through LAPTM4B-35 protein targeting.

Lysosome-associated protein transmembrane-4ß-35 is a novel independent prognostic factor in small cell lung cancer.

The lysosome-associated protein transmembrane-4ß-35 (LAPTM4B-35) protein has been indicated to be involved in solid tumors, while its role in small cell lung cancer (SCLC) remains unknown. The aim of this study is to investigate the LAPTM4B-35 protein expression and its clinical and prognostic role in SCLC patients. A total of 88 SCLC patients who underwent radical surgery between 2002 and 2010 were enrolled in the study. The level of messenger RNA (mRNA) and protein was detected from the fresh paired tumor specimens and adjacent normal tissues. The clinicopathological and survival data were collected. And the relationship between LAPTM4B-35 and clinicopathological features was analyzed. The prognostic value of LAPTM4B-35 for SCLC was investigated by univariate and multivariate analyses. The LAPTM4B-35 was overexpressed significantly in SCLC cancer tissues. The elevated protein expression was correlated strongly with clinical stage (p = 0.012) and tumor recurrence (p = 0.023). The 5-year overall survival and disease-free survival (DFS) were significantly worse in the patients with high LAPTM4B-35 level. Multivariate Cox analysis indicated that high LAPTM4B-35 expression was an independent prognostic factor for overall survival (OS) and DFS (p = 0.017 vs p = 0.011). LAPTM4B-35 overexpression was an independent factor in SCLC prognosis, which may be considered a potential useful marker in defining the SCLC prognosis.

LAPTM4B-35 protein is a weak tumor-associated antigen candidate.

Lysosome-associated protein transmembrane 4ß (LAPTM4B) is a gene that has been indicated to be involved in cancer. It is located at chromosome 8q22 and is composed of seven exons and six introns. LAPTM4B encodes two protein isoforms: LAPTM4B-35 and LAPTM4B-24. LAPTM4B-35 is markedly upregulated and LAPTM4B-24 is downregulated in several types of cancer. LAPTM4B-35 is 91 amino acids (N91) longer than LAPTM4B-24 at the N-terminus. In the present study, western blotting, enzyme-linked immunosorbent spot analysis and the B16F10-N91 tumor bearing-mice experiments were used to evaluate whether the overexpression of N91 indicates its potential as a candidate tumor-associated antigen. The results revealed that N91 was expressed in a wide range of normal mouse tissues and human peripheral blood mononuclear cells, with varying expression levels. The weak immunogenicity of N91 protein suggested it was a weak candidate antigen; however, the N91 protein was associated with cell proliferation.