Pleiotrophin promotes perineural invasion in pancreatic cancer

Perineural invasion(PNI)in pancreatic cancer is an important cause of local recurrence,but little is known about its mechanism.Pleiotrophin(PTN)is an important neurotrophic factor.It is of interest that our recent experimental data showed its involvement in PNI of pancreatic cancer.PTN strongly presents in the cytoplasm of pancreatic cancer cells,and high expression of PTN and its receptor may contribute to the high PNI of pancreatic cancer.Correspondingly,PNI is prone to happen in PTN-positive tumors.We thus hypothesize that,as a neurite growth-promoting factor,PTN may promote PNI in pancreatic cancer.PTN is released at the time of tumor cell necrosis,and binds with its highaffinity receptor,N-syndecan on pancreatic nerves,to promote neural growth in pancreatic cancer.Furthermore,neural destruction leads to a distorted neural homeostasis.Neurons and Schwann cells produce more N-syndecan in an effort to repair the pancreatic nerves.However,the abundance of N-syndecan attracts further PTN-positive cancer cells to the site of injury,creating a vicious cycle.Ultimately,increased PTN and N-syndecan levels,due to the continuous nerve injury,may promote cancer invasion and propagation along the neural structures.Therefore,it is meaningful to discuss the relationship between PTN/N-syndecan signaling and PNI in pancreatic cancer,which may lead to a better understanding of the mechanism of PNI in pancreatic cancer.     

胰腺癌神经浸润体外模型的构建与观察

目的 构建胰腺癌神经浸润的体外模型,并对胰腺癌细胞的神经浸润现象进行初步观察.方法 将胰腺癌Capan-2细胞与大鼠背根神经节(DRG)置于Matrigel基质胶中共培养,分别建立以观察细胞集落面积(模型1)和观察细胞迁移距离(模型2)为主的两种模型.倒置显微镜下观察神经突及细胞生长情况,利用图像分析软件Image pro plus对图片进行分析.结果 共培养对神经突起生长无显著影响,无论是共培养还是单培养,DRG神经突均向四周呈放射状生长.培养模型中,Capan-2细胞朝向DRG方向生长迁移,接着包绕神经突呈纺锤状,并继续向DRG爬行.在模型1的共培养组,Capan-2细胞第5天时细胞集落面积为(309.28±19.11) μm2,显著大于单培养模型的(208.57±7.94)μm2(P＜0.01).在模型2的共培养组,Capan-2细胞第5天时向DRG方向迁移了(284.1±12.9)μm,而空白基质胶一侧,细胞迁移距离＜150 μm,二者差异具有统计学意义(P＜0.01).结论 本实验成功构建了胰腺癌神经浸润体外模型,为后续研究打下了良好的实验基础.     

Nerve growth factor regulates CD133 function to promote tumor cell migration and invasion via activating ERK1/2 signaling in pancreatic cancer

BackgroundPerineural invasion (PNI) is extremely high frequency among the various metastatic routes in pancreatic cancer. Nerve growth factor, secreted by astroglial cells, exerts effects on tumor invasion in some cancer cells, but its function on migration and invasion in pancreatic cancer is still unclear. In the present study, we determined the effects of NGF on modulating tumor cell metastatic potential and invasion activity and explored its mechanisms in pancreatic cancer.MethodsNGF and CD133 expression were detected in tumor tissues using immunohistochemical analysis and Western blotting analysis. The effects of NGF on the regulation of CD133 expression and the promotion of cancer migration and invasion were investigated using wound healing and matrigel transwell assay. A related mechanism that NGF regulates CD133's function via activating ERK1/2 signaling also was observed.ResultsNGF/CD133 is overexpressed in human pancreatic cancer and promotes the migration and invasion of human pancreatic cancer cells through the activation of the ERK/CD133 signaling cascade. NGF/ERK signaling modulates the cancer cell EMT process, migration and invasion through the regulation of CD133 expression and its subcellular localization.ConclusionsNGF/CD133 signaling initiated the migration and invasion of pancreatic cancer cells. NGF/CD133 might be an effective and potent therapeutic target for pancreatic cancer metastasis, particularly in PNI.     

Midkine在胰腺癌组织中的表达及临床意义

目的 探讨人胰腺癌组织中Midkine（MK）的表达及其与肿瘤细胞增殖的关系和意义。方法 免疫组织化学SP法检测49例胰腺癌、13例慢性胰腺炎及15例正常胰腺组织中MK和Ki67蛋白的表达。结果胰腺癌组织中MK和Ki67均高表达，阳性率分别为77．1％（35／49）、81．6％（40／49），二者的表达都与肿瘤组织学分级、临床分期和淋巴结转移有关（P〈0．05）。胰腺癌组织中Ki67表达显著高于慢性胰腺炎（3／13，P〈0．01）和正常胰腺组织（0／15，P〈0．01）。慢性胰腺炎和正常胰腺组织中未见MK阳性表达。胰腺癌组织中MK蛋白的表达与Ki67的表达呈正相关（r-0．4，P〈0．05）。结论 检测MK蛋白对于胰腺癌的诊断及与慢性胰腺炎的鉴别诊断具有参考价值。MK在胰腺癌中高表达可能与肿瘤的发生发展及细胞增殖密切相关。     

与胰腺癌神经浸润相关的临床因素分析

目的 观察胰腺癌的神经浸润状况,分析与其相关的临床因素.方法 回顾性分析73例胰腺癌患者的神经浸润状况,分析神经浸润与肿瘤临床病理特征及患者生存率之间的关系.结果 73例中38例(52.1％)有神经浸润,其中6例(15.8％)为单纯胰内神经浸润,32例(84.2％)为胰内、胰外神经均浸润.神经浸润与患者性别、年龄及肿瘤病理类型、分化程度、大小、淋巴结转移均无关(P值均＞0.05),而与腹痛、脉管浸润、肿瘤组织表皮生长因子受体(EGFR)及血管内皮生长因子(VEGF)表达均显著相关(P值均＜0.01).有神经浸润患者的中位生存时间为8个月,显著短于无神经浸润患者的13个月(x2=4.69,P=0.030).结论 胰腺癌的神经浸润发生率较高,可引起明显腹痛,其与脉管浸润及肿瘤组织EGFR和VEGF表达相关,是影响胰腺癌患者术后生存率的因素之一.     

胰腺癌伴神经浸润动物模型的建立

目的 建立胰腺癌伴神经浸润的动物模型.方法 32只裸鼠分为4组,每组8只.将人胰腺癌细胞SW1990、CAPAN-2、PANC1分别注射于裸鼠的坐骨神经周围,以不做任何处理的裸鼠作为对照组.观察术后裸鼠体质量变化、成瘤情况、成瘤时间、造模成功率等指标.结果 对照组裸鼠体质量增加,各癌细胞注射组裸鼠成瘤后体质量明显下降,甚至呈现恶液质,成瘤侧肢体活动受限.CAPAN-2注射组及SW1990注射组的8只裸鼠最终均成瘤,PANC1注射组只有5只成瘤.以肿瘤最长径长至约0.8 cm为时间截点,CAPAN-2注射组、PANC1注射组、SW1990注射组裸鼠的成瘤时间分别为（49.8±5.0）、（56.6±2.4）、（25.4±3.0）d.SW1990注射组成瘤时间最短,PANC1注射组成瘤时间最长,3组间成瘤时间的差异具有统计学意义（F=73.51,P＜0.01）.CAPAN-2注射组、PANC1注射组、SW1990注射组裸鼠神经浸润造模成功率分别为87.5％ （7/8）、20.0％（1/5）、50.0％ （4/8）.结论 成功建立了胰腺癌伴神经浸润动物模型,但不同的胰腺癌细胞系建立的动物模型具有不同的特点.     

花姜酮对胰腺癌细胞迁移和侵袭的影响及机制的研究

目的:探讨花姜酮(zerumbone)对人胰腺癌细胞株BxPC-3和Panc-1迁移和侵袭的影响及其相关基因的表达。方法:以BxPC-3和Panc-1为研究对象,通过四甲基偶氮唑盐(MTT)实验选择毒性较小的花姜酮浓度,用细胞划痕实验、Transwell小室迁移和侵袭实验检测药物对细胞迁移和侵袭能力的影响,蛋白质印迹(Western blot)检测各细胞组CXC族趋化因子受体4(CXCR4)、促分裂原活化蛋白酶激酶1/2(MEK1/2)、磷酸化MEK 1/2(p-MEK1/2)、细胞外调节激酶1/2(ERK1/2)、磷酸化ERK1/2(p-ERK1/2)蛋白表达变化。结果:花姜酮对BxPC-3和Panc-1细胞的生长均有抑制作用,且随着药物浓度的增高和作用时间的延长而增强(P<0.05)。低浓度药物作用后BxPC-3和Panc-1的划痕迁移率都低于对照组(P<0.05)。与对照组相比,Transwell迁移和侵袭实验中BxPC-3和Panc-1用药组的平均穿膜细胞数均减少(P     

胰腺癌513例神经浸润的病理分析

目的 探讨胰腺癌神经浸润的特征及其与其他临床病理参数之间的关系.方法 光镜下观察491例胰腺导管腺癌、22例其他胰腺恶性肿瘤、41例胰腺良性病变和21例慢性胰腺炎组织中的神经浸润状况,分析其与其他病理学指标的相关性.结果 胰腺导管腺癌的神经浸润率为74%,显著高于其他类型恶性肿瘤的23%(P＜0.01).导管腺癌癌细胞通常穿越外周神经中膜到达内部的神经纤维束,有的甚至横断整根神经纤维.但神经浸润与导管腺癌的分化程度无关.52%的胰腺导管腺癌癌旁组织呈慢性炎症改变,且程度严重,远高于其他类型胰腺癌(14%)及胰腺良性病变(15%)的慢性炎症发生率(P＜0.01).胰腺导管腺癌淋巴细胞浸润神经的发生率为65%,远高于其他恶性肿瘤的36%和胰腺良性病变的22%(P＜0.01).胰腺导管腺癌的神经浸润与癌旁慢性胰腺炎症以及淋巴细胞浸润神经均相关,但与淋巴结转移无关.结论 神经浸润是胰腺导管腺癌特征性的生物学行为之一.     

Clinicopathological study of pancreatic carcinoma with particular reference to the invasion of the extrapancreatic neural plexus

A clinicopathological study of 44 ductal carcinomas of the head of the pancreas revealed 39 with retroperitoneal invasion, of which 27 showed extrapancreatic plexus involvements. The second portion of the plexus pancreaticus capitalis was the most frequent site of invasion. A statistically significant correlation was found between neural invasion in the pancreatic tissue and plexus invasion, but no clear correlation was found between plexus invasion and lymphatic invasion or tumor size. Even small-sized tumors (t₁) showed plexus invasion. The cases with plexus invasion had a statistically higher incidence of lymph-node involvement around the superior mesenteric artery than those without plexus invasion. These results indicate that complete dissection of extrapancreatic plexus around the superior mesenteric artery, including lymph nodes and soft tissue, could prolong the survival of patients with ductal carcinoma of the pancreas, even in cases of small-sized carcinomas.     

The complementary role of lymphovascular invasion and perineural invasion in the TNM staging process of rectal cancer

The aim of this study is to clarify the association between lymphovascular invasion (LVI) and/or perineural invasion (PNI) and the clinical characteristics and prognostic importance of rectal cancer, to provide a basis for early adjuvant treatment of rectal cancer. We retrospectively analyzed patients diagnosed with rectal cancer. This study involved rectal cancer tissue samples were obtained by surgical methods. Data on histological form, tumor classification, tumor size, gross growth pattern, blood and lymphatic vessel invasion, and PNI of the slice by HE staining were obtained from pathological examination. Immunohistochemical analysis of tissue samples was performed to determine p53 and EGFR expressions. There were 330 rectal cancer patients included in the study. LVI and/or PNI can be used as a high-risk factor for the prognosis of rectal cancer, predict prognostic survival, and guide adjuvant therapy. The detection rates of LVI and PNI were 32.1% and 16.1%. Differentiation grade, Union for International Cancer Control staging, tumor-lymph node-metastasis staging are significantly related to LVI or PNI. Multivariate logistic regression analysis shows that poor differentiation and N ≥ 1 can be used as independent risk factors and predictive factors for LVI. At the same time, poor differentiation and T > 3 is an independent risk factor for PNI. Only poor differentiation is the risk factor for poor prognosis in Cox risk regression analysis. In addition, the simultaneous occurrence of LVI and PNI is an independent prognostic factor.     

Preliminary study correlating CX3CL1/CX3CR1 expression with gastric carcinoma and gastric carcinoma perineural invasion

AIM:To study the relationship between the CX3CL1chemokine,its receptor CX3CR1,and gastric carcinoma/gastric carcinoma perineural invasion(PNI).METHODS:Thirty cases of gastric carcinoma were surgically resected(radical resection or palliative resection)between February 2012 and July 2012.Tumour and tumour-adjacent tissues were evaluated for the presence of CX3CL1(ELISA)and CX3CR1(immunohistochemistry and Western blotting)in an effort to analyse the relationship between CX3CL1/CX3CR1 and gastric carcinoma/gastric carcinoma PNI.RESULTS:Of these 30 cases,14 were PNI-positive(46.7%).No significant differences in CX3CL and CX3CR1 expression in tumour-adjacent tissues were found between the PNI positive and negative groups.Expression levels of CX3CL and CX3CR1 in tumour tissues were significantly higher than those in adjacent tissues(P<0.01),and were significantly higher in tumour tissues from the PNI-positive group compared to the PNI-negative group(P<0.01).CONCLUSION:CX3CL1/CX3CR1 expression may be associated with the occurrence and development of gastric carcinoma as well as gastric carcinoma PNI.     

胰星状细胞对胰腺癌细胞株Patu8988侵袭和转移的影响

目的:探讨永生化人胰星状细胞(IPSC)对胰腺癌细胞株Patu8988侵袭和转移的影响.方法:制备IPSC上清,用IPSC上清和Patu8988共培养,以单独培养的Patu8988为对照,比较实验组与对照组细胞的增殖、黏附能力.侵袭、迁移能力,克隆形成以及抵抗H2O2诱导的Patu8988凋亡能力.结果:与单独培养的胰腺癌细胞株Patu8988相比,IPSC上清对胰腺癌细胞株Patu8988细胞的增殖、黏附、迁移、侵袭能力及克隆形成能力有促进作用(均p<0.05),并能抑制H2O2诱导的Patu8988的凋亡(P<0.05).结论:胰星状细胞可能通过增强胰腺癌细胞株Patu8988的增殖、黏附、迁移、侵袭能力及克隆形成能力,并抑制其的凋亡,在胰腺癌的发展和转移中起重要作用.     

MLLT10 promotes tumor migration,invasion, and metastasis in human colorectal cancer

Objectives: Colorectal cancer (CRC), one of the most aggressive gastrointestinal malignancies, is a frequently diagnosed life-threatening cancer worldwide. Most CRC patients have poor prognosis mainly because of frequent metastasis and recurrence. Thus, it is crucial to find out some new biomarkers and to show deeper insights into the mechanisms of CRC. MLLT10, Myeloid/lymphoid or mixed-lineage leukemia translocated to 10, also known as AF10, a recurrent MLL partner. In this study, we found that MLLT10 promotes CRC tumor invasion and metastasis both in vitro and in vivo.

Methods: Here, the expression of MLLT10 was evaluated by immunohistochemistry. Then, the plasmid and lentivirus particles for MLLT10 overexpression or knockdown were designed and constructed into SW620 and HT29 cells. Finally, cell proliferation assay, cell adhesion assay, transwell migration, and invasion assay were used to detect the migration and invasion ability of MLLT10 in CRC cells. A tail vein injection assay was employed to evaluate the role of MLLT10 in tumor metastases.

Results: MLLT10 expression was significantly higher in CRC tissues than in noncancerous tissues and was associated with some clinicopathological factors. In vitro, the overexpression of MLLT10 promoted CRC cell migration and invasion, while after MLLT10 was knocked down, the opposite results were observed. Furthermore, we used animal metastasis models to detect the function of MLLT10 in vivo, the results are same with the outcomes in vitro. In lung metastasis sites, the knockdown of MLLT10 in SW620 cells significantly inhibited Vimentin expression, whereas the E-Cadherin was increased.

Conclusions: These results indicate that MLLT10 regulates the metastasis of CRC cells via EMT.     

miR-551b-3p在胰腺癌细胞和组织中的表达及其临床意义

目的研究miR-551b-3p在胰腺癌细胞和组织中的表达,探讨其在胰腺癌发生发展中的临床意义。方法利用实时定量聚合酶链反应(RT-PCR)技术分别检测4种胰腺癌细胞和正常胰腺导管上皮细胞,以及76对胰腺癌组织和癌旁组织中miR-551b-3p的表达水平,并对miR-551b-3p的表达与胰腺癌患者临床病理学特征进行分析。结果 miR-551b-3p在PANC-1、Aspc-1、SW1990和Miapaca-2四种胰腺癌细胞中的相对表达量分别为(0.125±0.012)、(0.179±0.005)、(0.672±0.025)、(0.577±0.019),低于其在正常胰腺导管上皮细胞HPDE6C-7中的相对表达量,差异有统计学意义(P0.05)。在胰腺癌组织和癌旁组织中miR-551b-3p的ΔCt值分别为(7.254±0.112)和(3.993±0.098),差异有统计学意义(P0.001)。TNM分期Ⅰ、Ⅱ、Ⅲ和Ⅳ期胰腺癌组织中miR-551b-3p的ΔCt值分别为(4.343±0.032)、(5.325±0.112)、(6.987±0.098)和(9.132±0.212),差异有统计学意义(P0.001)。有淋巴结转移和无淋巴结转移患者的胰腺癌组织中miR-551b-3p的ΔCt值分别为(8.492±0.021)和(6.676±0.103),差异有统计学意义(P=0.012)。高、中、低分化胰腺癌组织中miR-551b-3p的ΔCt值分别为(5.349±0.092)、(6.129±0.112)、(8.454±0.065),差异有统计学意义(P=0.002)。结论 miR-551b-3p的表达与胰腺癌的分化程度、淋巴结转移以及TNM分期相关。miR-551b-3p可能是提示胰腺癌临床进展的潜在标志物。     

Effects of perineural invasion in prostate needle biopsy on tumor grade and biochemical recurrence rates after radical prostatectomy

To predict local invasive disease before retropubic radical prostatectomy (RRP), the correlation of perineural invasion (PNI) on prostate needle biopsy (PNB) and RRP pathology data and the effect of PNI on biochemical recurrence (BR) were researched. For patients with RRP performed between 2005 and 2014, predictive and pathologic prognostic factors were assessed. Initially all and D'Amico intermediate-risk group patients were comparatively assessed in terms of being T2 or T3 stage on RRP pathology, positive or negative for PNI presence on PNB and positive or negative BR situation. Additionally the effect of PNI presence on recurrence-free survival (RFS) rate was investigated. When all patients are investigated, multivariate analysis observed that in T3 patients PSA, PNB Gleason score (GS) and tumor percentage were significantly higher; in PNI positive patients PNB GS, core number and tumor percentage were significantly higher and in BR positive patients PNB PNI positivity and core number were significantly higher compared to T2, PNI negative and BR negative patients, separately (p < 0.05). When D'Amico intermediate-risk patients are evaluated, for T3 patients PSA and PNB tumor percentage; for PNI positive patients PNB core number and tumor percentage; and for BR positive patients PNB PNI positivity were significantly higher compared to T2, PNI negative and BR negative patients, separately (p < 0.05). Mean RFS in the whole patient group was 56.4 ± 4.2 months for PNI positive and 96.1 ± 5.7 months for negative groups. In the intermediate-risk group, mean RFS was 53.7 ± 5.1 months for PNI positive and 100.3 ± 7.7 months for negative groups (p < 0.001). PNI positivity on PNB was shown to be an important predictive factor for increased T3 disease and BR rates and reduced RFS.     

Peroxisome proliferator-activated receptor gamma ligand inhibits cell growth and invasion of human pancreatic cancer cells

Summary Background. Peroxisome proliferator-activated receptor γ (PPARγ) is expressed in certain human cancers; ligand-induced PPARγ activation can result in growth inhibition and differentiation in these cells. However, the precise mechanism for the antiproliferative effect of PPARγ ligands is not entirely known. Aim of Study. The purpose of this study was to examine the effect of PPARγ ligands on pancreatic cancer cell growth and invasiveness. Methods. The effect of two PPARγligands, 15 deoxy-Δ^12,14prostaglandin J₂ (15d-PGJ₂) and ciglitazone, on the growth of four human pancreatic cancer cell lines (BxPC-3, MIA PaCa-2, Panc-1, and L3.6) was assessed. Expression of cell-cycle and apoptotic-related proteins was measured. Finally, the effect of 15d-PGJ₂ on pancreatic cancer cell invasiveness and matrix metalloproteinase expression was determined. Results. Both 15d-PGJ₂ and ciglitazone inhibited the growth of all four pancreatic cancer cell lines in a dose- and time-dependent fashion. Treatment of BxPC-3 cells with 15d-PGJ₂ resulted in a time-dependent decrease in cyclin D1 expression associated with a concomitant induction of p21^waf1 and p27^kip1. In addition, 15d-PGJ₂ treatment induced apoptosis through activation of caspase-8, -9, and -3. Moreover, pancreatic cancer cell invasiveness was significantly suppressed after treatment with a nontoxic dose of 15d-PGJ₂, which was associated with a reduction of MMP-2 and MMP-9 protein levels and activity. Conclusion. These results demonstrate that PPARγ ligands have the dual advantage of inhibiting pancreatic cancer cell growth while reducing the invasiveness of the tumor cells, suggesting a potential role for these agents in the adjuvant treatment of pancreatic cancer.