Baoquan Liu

Lymphangiogenesis and Its Relationship With Lymphatic Metastasis and Prognosis in Malignant Melanoma

Regional lymph node metastasis is one of the important indicators of cutaneous malignant melanoma. Newly formed lymphatic vessels are considered to provide a route whereby tumor cells can migrate to the lymph nodes. Both vascular endothelial growth factors (VEGF) ‐C and ‐D have been confirmed to participate in tumor lymphangiogenesis, but the prognostic significance of VEGF‐C, VEGF‐D, and lymphangiogenesis in cutaneous malignant melanoma remains controversial. To clarify the effects of these factors and to evaluate the relationships between lymphangiogenesis, lymph node metastasis, and prognosis in patients with malignant melanoma, the expressions of VEGF‐C, VEGF‐D, and their receptor (VEGFR) ‐3 were detected by immunohistochemistry and reverse transcriptase‐polymerase chain reaction. The expressions of both VEGF‐C and VEGF‐D proteins were concomitantly detected in the cytoplasm of the malignant cells. VEGF‐C and VEGF‐D expressions were associated with VEGFR‐3 expression and were significantly correlated with both peritumoral lymphangiogenesis and lymph node metastasis. The incidence of peritumoral lymphatic vessels was significantly higher in lymph node metastatic melanomas than that in nonmetastatic melanomas. Univariate and multivariate analyses indicated that VEGF‐C and VEGF‐D were independent prognostic factors for overall survival and disease‐free survival in patients with malignant melanoma. This study suggests that both VEGF‐C and VEGF‐D are involved in peritumoral lymphangiogenesis and lymphatic metastasis. VEGF‐C and VEGF‐D expression may be clinically useful indicators for prognostic evaluation in patients with cutaneous malignant melanoma. Anat Rec, 2008.

Vascular Endothelial Growth Factor D and Intratumoral Lymphatics as Independent Prognostic Factors in Epithelial Ovarian Carcinoma

Lymph node metastasis is an important prognostic indicator for disease progression and is crucial for therapeutic strategies of epithelial ovarian carcinoma. Vascular endothelial growth factor (VEGF)‐D has been confirmed to have potent lymphangiogenic function in experimental models, but the role in the progression of human ovarian carcinoma remains presently controversial. The purpose of this study was to investigate the prognostic significance of VEGF‐D and the presence of intratumoral lymphatics in patients with epithelial ovarian carcinoma. The VEGF‐D expression was evaluated by immunohistochemistry in 78 specimens of epithelial ovarian carcinoma and tumoral lymphatic vessels were measured by D2‐40. The expression of VEGF‐D protein was detected in the cytoplasm of the tumor cells and in stroma occasionally. The high expression of VEGF‐D was closely associated with the FIGO stage, intratumoral lymphatic vessels, tumoral lymphatic invasion, and lymph node metastasis as well as a shorter overall survival. Univariate and multivariate analysis indicated that VEGF‐D, intratumoral lymphatics, and lymphatic invasion were independent prognostic factors for overall survival and disease‐free survival in patients with epithelial ovarian carcinoma. We conclude that VEGF‐D plays an essential role in tumoral lymphangiogenesis and lymphatic spread, VEGF‐D expression, and the intratumoral lymphatics may be clinically useful indicators for prognostic evaluation in patients with epithelial ovarian carcinoma. Anat Rec, 2009.

SCARA5 plays a critical role in the progression and metastasis of breast cancer by inactivating the ERK1/2, STAT3, and AKT signaling pathways

Scavenger receptor class A member 5 (SCARA5) is a candidate anti-oncogene in several malignancies. However, whether SCARA5 is a suppressor gene in breast cancer and its role in breast cancer cell growth and metastasis remain to be determined. Here, we investigated the biological functions of SCARA5 in the progression and metastasis of breast cancer and explored the underlying mechanisms. A total of 65 breast cancer patients and three cell lines (ZR-75-30, MCF-7, and MDA-MB-231) were analyzed in the study. RT-qPCR, western blotting, and immunohistochemistry were used to detect mRNA and protein expression, and lymphatic vessel density (LVD) and microvessel density (MVD). MTT, colony formation, TUNEL assays, invasion assays and Transwell assays, and flow cytometric analyses were used to evaluate the effect of SCARA5 on breast cancer cells. SCARA5 was significantly downregulated in breast cancer tissues and cells and significantly correlated with tumor size, histological grade, lymph node metastasis, pTNM stage, VEGF-A, VEGF-C, LVD, and MVD. SCARA5 overexpression significantly suppressed cell proliferation, colony formation, invasion, and migration, and induced G0/G1 arrest and apoptosis of ZR-75-30 cells. SCARA5 decreased the phosphorylation of ERK1/2, AKT, and STAT3, and downregulated downstream signaling effectors, including MMP-2, 3, and 9, VEGF-A, VEGF-C, Bax, Cyclin B1, Cyclin D1, and Cyclin E1, and upregulated E-cadherin, Bcl-2, and caspase 3. SCARA5 is associated with multiple signaling pathways and plays a critical role in the progression and metastasis of breast cancer. The present results provide the first evidence that SCARA5 inhibits lymphangiogenesis by downregulating VEGF-C, thereby inhibiting breast cancer lymphatic metastasis.

Serum miR-21 and miR-125b as markers predicting neoadjuvant chemotherapy response and prognosis in stage II/III breast cancer

The predictive value of serum miRNAs (ser-miRNA) for the response to neoadjuvant chemotherapy (NCT) and the prognosis of breast cancer patients were investigated in the current study. The study included 118 stage II/III breast cancer patients and 30 healthy adult women. Peripheral blood was drawn from participants before the start (baseline [BL]), at the end of the second cycle (first evaluation during NCT [FEN]), and at the end of NCT (second evaluation during NCT [SEN]). The expression of ser-miRNAs was examined by quantitative reverse-transcription polymerase chain reaction (qRT-PCR), and their association with chemotherapy response and prognosis was analyzed. MiR-19a, miR-21, miR-125b, miR-155, miR-205, and miR-373 were significantly up-regulated in the serum of breast cancer patients at BL, miR-451 was significantly down-regulated, and miR-122 was unchanged compared with the levels in healthy women. The expression of ser-miR-125b and the changes of ser-miR-21 expression during NCT were associated with chemotherapy response and disease-free survival (DFS). In chemotherapy responders, ser-miR-125b expression was lower than that of non-responders at BL, FEN, and SEN, and ser-miR-21 levels decreased from BL to FEN and from BL to SEN. Survival analysis showed that patients with lower ser-miR-125b expression at BL, FEN, and SEN had favorable DFS, and those with decreased ser-miR-21 expression from BL to FEN and from BL to SEN had better DFS. In conclusion, ser-miR-21 and ser-miR-125b were identified as novel, noninvasive predictive markers for NCT response and prognosis in breast cancer.

A novel seven-long non-coding RNA signature predicts survival in early stage lung adenocarcinoma

Increasing evidence has revealed the significant association between dysregulated lncRNA expression and cancers. The prognostic value of lncRNAs in predicting the risk of disease recurrence and identifying high-risk subgroup of early stage lung adenocarcinoma (LUAD) is still unclear. In this study, we analyzed lncRNA expression profiles of 415 early-stage LUAD patients from Gene Expression Omnibus and identified a novel seven-lncRNA signature that was significantly associated with survival in patients with early-stage LUAD (HR = 2.718, CI = 2.054–3.597, p < 0.001). Based on the seven-lncRNA signature, we constructed a risk score model which is able to classify patients of training dataset into the high-risk group and the low-risk group with significantly different clinical outcome (p < 0.001). The robustness of the seven-lncRNA signature was successfully validated through application in other two independent patient datasets. Furthermore, the prognostic value of seven-lncRNA signature was independent of other clinicopathological factors including age, gender, stage and smoking status. Functional analysis suggested that the seven-lncRNA signature may be involved in a variety of biological pathways including cell cycle, ECM-receptor interaction, Focal adhesion and p53 signaling pathway. Taken together, our study not only provides insights into the lncRNA association with LUAD, but also provide alternative molecular markers in prognosis prediction for early-stage LUAD patients.

Integrin α4 Induces Lymphangiogenesis and Metastasis via Upregulation of VEGF-C in Human Colon Cancer

Vascular endothelial growth factor‐C (VEGF‐C) is a key regulator in lymphangiogenesis, and is overexpressed in various malignancies. Integrin α4β1, a new member of the VEGF‐C/VEGF receptor pathway, was found to be overexpressed in melanoma tumors. However, little is known regarding the potential role of integrin α4β1 in lymphangiogenesis and other solid tumors. The aim of this study was to investigate the expression patterns of integrin α4 and VEGF‐C in relation to lymphangiogenesis and clinicopathological parameters in human colon cancer. The expression of integrin α4, VEGF‐C, and VEGFR‐3 was assessed in 71 human colon cancer tissues and 30 paracancerous normal tissues by immunohistochemical staining. Lymphatic microvessel density (LMVD) was measured after D2‐40‐labeling, and the correlations among different factors were statistically analyzed. The expression of integrin α4, VEGF‐C, VEGFR‐3, and LMVD was higher in colon cancer tissues compared with the normal paracancerous colon tissues. There was a positive correlation between the expression of integrin α4 and VEGF‐C. Integrin α4 and VEGF‐C were significantly associated with the clinicopathological parameters (LMVD, Dukes stage, and lymph node metastasis). Kaplan–Meier analyses indicated that patients with high integrin α4 or VEGF‐C expression had significantly shorter overall survival and tumor‐free survival time. Multivariate analyses suggested that integrin α4 and VEGF‐C may serve as independent prognostic factors for human colon cancer. Both integrin α4 and VEGF‐C are involved in lymphangiogenesis and lymphatic metastasis. Our results demonstrated that integrin α4 is a novel prognostic indicator for human colon cancer. Anat Rec, 299:741–747, 2016.

Pancreatitis‐Associated Protein Is Related Closely to Neoplastic Proliferative Activity in Patients with Colorectal Carcinoma

Pancreatitis‐associated protein (PAP) is a secretory protein that is not only expressed during acute pancreatitis but also in pancreatic adenocarcinoma, gastric carcinoma, hepatocellular carcinoma, and colorectal carcinoma. Expression in carcinoma might be another characteristic of PAP. The aim of our study was to assess, in 27 patients undergoing surgery for colorectal carcinoma, the expression of the PAP mRNA and to evaluate its association with DNA ploidy and proliferative activity (S‐phase fraction, SPF) by reverse transcriptase‐polymerase chain reaction (RT‐PCR) and flow cytometric analysis (FCM). PAP mRNA was expressed in 29.6% (8 of 27) of the patients with colorectal carcinoma. DNA aneuploid and high SPF were found in 87.5% (7 of 8) of patients with PAP mRNA positive colorectal carcinoma. The serum PAP level was significantly elevated in patients with colorectal carcinoma when compared with the healthy subjects. Twelve of the 27 patients with colorectal carcinoma had high serum PAP concentrations (>25 ng/mL) and the mean SPF was 17.82% ± 8.02%, which was significantly higher compared with the normal colorectal tissue group (7.33% ± 3.18%, P < 0.05). The mean serum PAP concentration of DNA aneuploidy colorectal carcinomas was 46.67 ± 17.58 ng/mL, which was significantly different when compared with the DNA diploidy group (19.18 ± 8.89 ng/mL, P < 0.05). PAP mRNA expression and serum PAP levels are closely related to neoplastic proliferative activity in patients with colorectal carcinoma. No significant differences are observed between PAP mRNA expression and clinicopathologic parameters (P > 0.05). Anat Rec, 2009.

Changes of serum miR34a expression during neoadjuvant chemotherapy predict the treatment response and prognosis in stage II/III breast cancer

OBJECTIVE To investigate the predictive value of serum miR34a (ser-miR34a) expression for the neoadjuvant chemotherapy (NACT) response and prognosis in breast cancer patients. METHODS This study included 86 diagnosed stage II/III breast cancer patients and 20 healthy volunteers. Peripheral blood from every participant was collected before the start, at the end of the second cycle, and at the end of NACT. The expression of ser-miR34a was examined by qRT-PCR and its association with the chemotherapy response and prognosis was analyzed. RESULTS The expression of ser-miR34a in breast cancer patients before NACT was significant higher than that of healthy volunteers. During the NACT, the changes in ser-miR34a expression were significantly associated with treatment response and disease-free survival (DFS). In responding patients, ser-miR34a levels at the end of the second cycle and at the end of NACT were significantly lower than before NACT (P=0.016 and P=0.002, respectively), and in non-responding patients, the changes were insignificant. Survival analyses showed that the patients with decreased ser-miR34a expression from the end of the second cycle and the end of NACT to before NACT had improved DFS compared with that of the patients with increasing ser-miR34a expression (P<0.001 for both). Cox regression analyses showed that the changes of ser-miR34a expression were independent prognostic indicators. CONCLUSIONS Ser-miR34a is a novel, noninvasive predictive marker for NACT response and prognosis in breast cancer patients.

Smad4 inhibits VEGF-A and VEGF-C expressions via enhancing Smad3 phosphorylation in colon cancer

Smad4 is a critical factor in the TGF‐β pathway and is involved in tumor progression and metastasis, but the role of Smad4 in colon cancer cells is unclear. The aim of this study is to explore the effect and the underlying mechanism of Smad4 on the growth, migration and apoptosis of colon cancer cells as well as vascular endothelial growth factor (VEGF)‐A and VEGF‐C secreted by these cells. In this study, we showed that Smad4, VEGF‐A, and VEGF‐C are independent prognostic factors of colon cancer, and Smad4 expression was negatively correlated with VEGF‐A and ‐C in samples. We found that Smad4 mRNA and protein levels in colon cancer cells, particularly in HCT‐116 cells, were significantly lower than those in the human intestinal epithelial cell line (HIEC). Smad4 overexpression promoted tumor cell apoptosis, inhibited VEGF‐A and ‐C expression in vitro and in vivo, but had no effect on cell proliferation and migration. Tail vein injection of the virus inhibited xenograft growth in nude mice. Importantly, we also demonstrated that Smad4 could increase the phosphorylation level of Smad3, but not Smad2, which may be one of the mechanisms underlying these effects of Smad4 in colon cancer. Therefore, Smad4 may be a new target for the treatment of colon cancer. Anat Rec, 300:1560–1569, 2017.

Association between VEGF-A, C and D expression and lymph node involvement in breast cancer: a meta-analysis

Background Metastasis is the primary cause of death in patients with breast cancer. Although VEGF-A, C and D are considered to be prime factors in lymph node metastasis in breast cancer, the published studies have conflicting conclusions. Methods To resolve this conflict, we conducted a meta-analysis of 37 studies (n = 5,001 patients) evaluating the correlation between VEGF-A, C and D immunohistochemical expression and lymph node metastasis (LNM). The meta-analysis included 22 studies of VEGF-A, 17 of VEGF-C, and 6 of VEGF-D. The relationships between VEGF-A, C and D and clinicopathological parameters were also examined. Results The results showed a significant association between VEGF-A or VEGF-C overexpression and LNM (risk ratio [RR] = 1.28 [95% CI 1.04-1.58], p = 0.02; and RR = 1.36 [95% CI 1.07-1.72], p = 0.01, respectively). Subgroup evaluation showed a significant association between VEGF-A, C and D overexpression and LNM when analyses were limited to Asian patients (RR = 1.78 [95% CI 1.28-2.46], p = 0.0005; RR = 1.38 [95% CI 1.04-1.84], p = 0.03, and RR = 2.62 [95% CI 1.35-5.09], p = 0.004, respectively). VEGF-A overexpression was significantly associated with lymph vessel invasion (RR = 1.86 [95% CI 1.33-2.60], p = 0.0003). Overexpression of VEGF-C or VEGF-D was significantly associated with HER-2 positivity (RR = 1.30 [95% CI 1.06-1.59], p = 0.01; and RR = 1.75 [95% CI 1.01-3.03], p = 0.05, respectively). Conclusions With some limitations, our meta-analysis indicated that VEGF-A and C could predict LNM in patients with breast cancer, particularly Asian patients.