Tumor‐associated macrophages induce lymphangiogenesis in cervical cancer via interaction with tumor cells

Our studies were conducted to investigate the clinical and functional significance of tumor‐associated macrophages (TAMs) in cervical tumor lymphatic metastasis. We found that the increase in macrophages in tumor stroma is significantly associated with lymphatic metastasis (p = 0.017), through performing immunohistochemical staining in 111 cervical samples (55 invasive squamous carcinomas of uterine cervix, 27 cervical intraepithelial neoplasms III, and 29 normal cervix). The human lymphatic endothelial cells (HLEC), which were cultured in conditioned medium of cervical cancer cell‐macrophage coculture, formed more tube‐like structures in vitro, when compared with those in conditioned mediums of LEC, normal cervical epithelium, single macrophage, and single cervical cancer cell (all p < 0.001). The mRNA expressions of IL‐1β and IL‐8 in cervical cancer cells cocultured with macrophages were increased, compared with those in cervical cancer cell cultured alone (p_IL‐1β < 0.05 and p_IL‐8 < 0.01). Meanwhile, the mRNA expression of VEGF‐C and VEGF‐A was increased in macrophages cocultured with cervical cancer cells, compared with the expression in those macrophages cultured alone (both p < 0.05). Taken together, the results suggest that TAMs promote lymphangiogenesis mainly through interaction with surrounding cervical cancer cells.     

Prognostic Value of Lymphangiogenesis Determinants in Luminal and Non-luminal Breast Carcinomas

Background: Breast carcinomas (BCs) are sub-classified according to the molecular characteristics into luminal andnon-luminal subtypes that clinically show different biological behavior, treatment and prognosis. BCs spread primarilythrough lymphatic vessels using cascade processes of lymphagiogenesis in which VEGF-C plays an important role duringlymph node metastasis. Prognostic value of VEGF-C in luminal and non-luminal BC is still unclear and has not beenstudied thoroughly to clarify and define prognosis and therapeutic monitoring. Aim: To define the prognostic value oflymphangiogenesis on survival rates of luminal and non-luminal subtypes BC. Materials and Methods: This studyapplied prospective cohort design, using 130 patients of invasive duct carcinoma of the breast, stage I-IIIA, fromSardjito General Hospital, Indonesia and subsequent longitudinal follow-up. Immunohistochemical staining wascarried out using anti-ER, -PR, -Her-2, VEGF-C, VEGFR-3 and D2-40 antibodies. The related clinicopathologiccharacteristics of BC patients and lymphangiogenesis determinants, including VEGF-C expression, were statisticallyanalyzed. Results: In non-luminal BC subtypes, VEGF-C expression (HR=0.04; 95% CI=0.01-0.41), lymph nodemetastasis (HR=0.14; 95% CI=0.04-0.55) and stage (HR=0.30; 95% CI= 0.02-0.76) were determined as independentprognostic factors on survival rates. However, the lymphangiogenesis determinants were not associated with the survivalrates of luminal BC subtypes. Conclusion: This study suggested that lymphangiogenesis affects survival rates ofnon-Luminal subtype rather than the luminal subtypes of BC.     

胰腺癌淋巴管生成及其与生物学行为关系

目的研究胰腺癌淋巴管生成的机制、分布特征及其与胰腺癌临床病理学的关系。方法应用免疫组化双标记方法检测42例胰腺癌和12例癌旁胰腺组织中血管内皮生长因子-C（VEGF—C）及肾小球足突细胞黏蛋白（Podoplanin）的表达。结果42例胰腺癌组织VEGF—C阳性率为61．9％，12例癌旁胰腺组织中VEGF—C阳性表达率为58．3％，两者差异无显著性（x^2=0．050，P〉0．05）。VEGF-C阳性率与胰腺癌的淋巴结转移有密切关系（x^2=4．822，P〈0．05），而与胰腺癌大小、组织学分级、神经浸润及临床病理分期无关（P〉0．05）。胰腺癌组织中可见明确的Podoplanin阳性淋巴管，其数目和分布具有明确的异质性。肿瘤边缘组织中淋巴管数最多，其次为肿瘤表浅部，而肿瘤中心区最少；在形态上，肿瘤边缘可以见到较多管腔扩张的淋巴管，而肿瘤中心及表浅淋巴管多为闭锁的条索状或点状。癌旁胰腺中Podoplanin阳性淋巴管分布在呈慢性炎症的胰腺组织周围，多数呈扩张状态。42例胰腺癌组织中LVD为7．67±1．25，12例癌旁胰腺组织中LVD为7．85±0．93，二者差别无显著性（t=0．639，P〉0．05）。胰腺癌组织中LVD与淋巴结转移及临床病理分期有关（t=7．076，6．803，P〈0．01），而与胰腺癌大小、组织学分级和神经浸润无关（P〉0．05）。胰腺癌组织中VEGF-C的表达与LVD间呈正相关性（r=0．509，P〈0．05）。结论胰腺癌及癌旁胰腺组织中VEGF—C高表达，促进淋巴管增生，可能是临床上胰腺癌早期发生淋巴结转移的重要机制之一。     

Changes in Lymphangiogenesis and Vascular Endothelial Growth Factor Expression by Neo‐Adjuvant Hormonal Therapy in Prostate Cancer Patients

BACKGROUND

The anti‐cancer mechanism of neo‐adjuvant hormonal therapy (NHT) is not well understood. Lymphangiogenesis plays an important role in cancer progression and is regulated by a complex mechanism that includes vascular endothelial growth factor (VEGF) signaling. However, there is little information regarding relationship between lymphangiogenesis and androgen deprivation. The aim of this study was to clarify changes in lymphangiogenesis and VEGF expression induced by androgen deprivation in prostate cancer in vivo and in vitro.

METHODS

Patients who had undergone a radical prostatectomy were enrolled in the study (NHT, n = 60 and non‐NHT, n = 64). Lymph vessels were identified by D2‐40 immunoreactivity and lymph vessel density and lymph vessel area (LVD and LVA, respectively) were measured from micrographs. The expression of VEGF‐A, ‐B, ‐C, and ‐D was evaluated by immunohistochemistry. The prognostic value of LVD and LVA for biochemical recurrence was also investigated.

RESULTS

Mean LVD ± SD was higher in the NHT than in the non‐NHT group (11.3 ± 3.0 vs. 7.1 ± 3.4 per high power field; P < 0.001). LVA was larger in the NHT than in the non‐NHT group (512.8 ± 174.9 vs. 202.7 ± 72.8 µm²; P < 0.001). VEGF‐A expression was lower whereas VEGF‐C and ‐D levels were higher in the NHT than in the non‐NHT group. VEGF‐B expression in specimens with NHT was lower than that in biopsy specimens at diagnosis. These results were confirmed by in vitro studies used androgen‐sensitive prostate cancer cell line. LVA was found to be an independent predictor of biochemical recurrence in patients who received NHT.

CONCLUSIONS

Our results demonstrate that NHT stimulates lymphangiogenesis via upregulation of VEGF‐C and ‐D, which may increase LVA and affect the outcome of prostate cancer patients. This findings were supported by in vitro data of prostate cancer cell. Prostate 77:255–262, 2017. © 2016 The Authors. The Prostate Published by Wiley Periodicals, Inc.     

心脏淋巴管新生及其与心血管疾病预后的关系

 心脏淋巴管的生理病理作用一直未引起人们的重视。但近年来,随着血管内皮生长因子受体-3(vascular endothelial growth factor receptor 3,VEGFR-3)、淋巴管内皮细胞透明质酸受体(lymphatic vessel endothelial hyaluronan receptor 1,LYVE-1)、果蝇同源基因转录因子(prospero homeobox protein 1,Prox-1)、肾小球足细胞膜黏蛋白(podoplanin)等淋巴管内皮细胞标志物的发现,心脏淋巴管的研究迅速发展。心脏淋巴管引流淋巴液,其功能异常可引起心肌水肿、动脉硬化、心律失常、间质纤维化等。深入认识心脏淋巴管的生理病理作用,对于治疗心肌梗死、改善心血管功能、预防心脏手术后并发症等有着重要意义。本文综述了心脏淋巴管的发生、分布以及与心血管疾病预后的关系。     

miR-377-5p通过下调VEGF-C对结肠癌的血管和淋巴管生成的影响研究

目的探讨miR-377-5p通过下调VEGF-C对结肠癌的血管和淋巴管生成的影响研究。方法该实验分为3组,miR-377-5p干扰组、miR-377-5p阴性对照组和miR-377-5p过表达组。采用实时荧光RT-PCR、Western blot、CCK-8、流式细胞、细胞划痕、Transwell、血管生成实验检测miR-377-5p m RNA和蛋白表达、细胞增殖、凋亡、迁移、侵袭、血管和淋巴管生成。结果与正常结肠组织对比,结肠癌组织中miR-377-5p m RNA和miR-377-5p蛋白的表达明显低于正常结肠组织(均P<0.05);与miR-377-5p干扰组相比,miR-377-5p阴性对照组和miR-377-5p过表达组中的m RNA及蛋白显著升高(均P<0.05),与miR-377-5p阴性对照组相比,miR-377-5p过表达组的m RNA及蛋白明显增加(P<0.05);与miR-377-5p干扰组相比,miR-377-5p阴性对照组和miR-377-5p过表达组的结肠癌细胞增殖能力明显降低(P<0.05),与miR-377-5p阴性对照组相比,miR-377-5p过表达组的结肠癌细胞增殖能力显著降低(P<0.05);与miR-377-5p干扰组相比,miR-377-5p阴性对照组和miR-377-5p过表达组的结肠癌细胞增殖凋亡明显升高(P<0.05),与miR-377-5p阴性对照组相比,miR-377-5p过表达组的结肠癌细胞凋亡能力显著升高(P<0.05);伤口愈合实验显示,miR-377-5p过表达组的结肠癌细胞的迁移能力显著低于miR-377-5p干扰组和miR-377-5p阴性对照组(P<0.05);miR-377-5p过表达组的结肠癌细胞的侵袭能力也明显低于miR-377-5p干扰组和miR-377-5p阴性对照组(P <0.05);miR-377-5p过表达组的VEGF-A、VEGF-C和P-Akt、VEGFR-3蛋白水平显著低于miR-377-5p干扰组和miR-377-5p阴性对照组(P<0.05);miR-377-5p过表达组的MVD(微血管密度)和LMVD(淋巴微血管密度)显著低于miR-377-5p干扰组和miR-377-5p阴性对照组(P<0.05)。结论 miR-377-5p在结肠癌中低表达,抑制了结肠癌细胞的增殖、细胞迁移和侵袭,促进了结肠癌细胞的凋亡能力,并直接靶向VEGF-C抑制肿瘤的血管和淋巴管生成。     

Diagnosis and management of lymphatic vascular disease

The lymphatic vasculature is comprised of a network of vessels that is essential both to fluid homeostasis and to the mediation of regional immune responses. In health, the lymphatic vasculature possesses the requisite transport capacity to accommodate the fluid load placed upon it. The most readily recognizable attribute of lymphatic vascular incompetence is the presence of the characteristic swelling of tissues, called lymphedema, which arises as a consequence of insufficient lymph transport. The diagnosis of lymphatic vascular disease relies heavily upon the physical examination. If the diagnosis remains in question, the presence of lymphatic vascular insufficiency can be ascertained through imaging, including indirect radionuclide lymphoscintigraphy. Beyond lymphoscintigraphy, clinically-relevant imaging modalities include magnetic resonance imaging and computerized axial tomography. The state-of-the-art therapeutic approach to lymphatic edema relies upon physiotherapeutic techniques. Complex decongestive physiotherapy is an empirically-derived, effective, multicomponent technique designed to reduce limb volume and maintain the health of the skin and supporting structures. The application of pharmacological therapies has been notably absent from the management strategies for lymphatic vascular insufficiency states. In general, drug-based approaches have been controversial at best. Surgical approaches to improve lymphatic flow through vascular reanastomosis have been, in large part, unsuccessful, but controlled liposuction affords lasting benefit in selected patients. In the future, specifically engineered molecular therapeutics may be designed to facilitate the controlled regrowth of damaged, dysfunctional, or obliterated lymphatic vasculature in order to circumvent or mitigate the vascular insufficiency that leads to edema and tissue destruction.