乳腺原发癌和相应淋巴结转移癌干细胞Wnt、Notch信号通路相关分子的比较

目的：探讨乳腺原发癌和相应淋巴结转移癌干细胞Wnt、Notch信号通路分子β-catenin、Cyclin D1和Notch1 mRNA表达及意义。方法选取乳腺浸润性导管癌30例,且均伴有淋巴结转移,制备乳腺癌单细胞悬液,通过免疫磁珠分选技术分别提取乳腺原发癌和相应淋巴结转移癌干细胞,采用实时荧光定量PCR技术检测两种癌干细胞Wnt、Notch信号通路关键分子β-catenin、Cyclin D1和Notch1 mRNA的表达。结果乳腺原发癌干细胞和相应淋巴结转移癌干细胞中β-catenin mRNA的表达差异无统计学意义(P>0.05);淋巴结转移癌干细胞中Cyclin D1和Notch1 mRNA的表达显著高于乳腺原发癌干细胞(P<0.01)。结论与乳腺原发癌干细胞相比,淋巴结转移癌干细胞中Cyclin D1和Notch1处于更高的活化状态,具有更高的侵袭和转移能力,有可能成为乳腺癌的新治疗靶点。     

鼻咽癌中Shh和Ptch1基因的表达及其临床意义

目的探讨鼻咽癌中Hedgehog信号通路成员Shh和Ptch1基因表达的关系及其意义。方法采用免疫组化En Vision两步法检测69例鼻咽癌及15例正常鼻咽黏膜中Shh、Ptch1蛋白表达,分析Shh、Ptch1蛋白表达水平与鼻咽癌临床病理特征的关系。结果鼻咽癌中Shh及Ptch1蛋白呈高表达分别为53.6%、37.68%,高于正常鼻咽黏膜组织(二者均为6.7%)(P均0.05)。Shh表达与肿瘤淋巴结转移、临床分期有关(P0.05);与肿瘤侵袭、远处转移、患者年龄及性别等均无关(P0.05);Ptch1蛋白表达水平与鼻咽癌临床病理特征的比较无统计学意义(P0.05);Shh与Ptch1蛋白表达呈正相关(r=0.456,P0.01)。结论 Shh和Ptch1蛋白高表达,可能参与鼻咽癌的发生、发展。     

Ptch和Smo蛋白在非小细胞肺癌中的表达及意义

目的观察非小细胞肺癌(non-small cell lung cancer,NSCLC)及癌旁组织中Ptch和Smo蛋白的表达,探讨Ptch和Smo表达与临床病理学的关系。方法应用免疫组织化学及免疫印迹技术检测NSCLC及癌旁组织中Ptch和Smo蛋白的表达。结果 Ptch和Smo蛋白在癌旁组织中不表达或低表达,但在NSCLC中表达明显增强。Ptch和Smo蛋白的表达与癌组织学分型无关,与癌组织分化程度和淋巴结转移密切相关,分化程度低、淋巴结转移阳性者,表达增强明显。结论过表达的Ptch和Smo蛋白可能参与NSCLC的发生、发展过程。     

Hedgehog信号传导通路在乳腺癌中表达增强

目的：研究乳腺癌细胞中,Hedgehog信号传导通路是否表现为持续的激活状态。方法：用免疫组化方法在64例乳癌标本中检测决定Hedgehog信号传导通路的组成部分,包括Shh,patched1和Gli1,SMO的表达。结果：在64例肿瘤样本中,强阳性表达Shh ,Ptch1和Glil,Smo的分别为64（100％）,61（95.3％）,62（96.9％）,和61（95.3％）,相反,各自对应相邻的正常乳腺上皮在可探测到的水平上不表达或低表达的这些蛋白,乳腺癌中的Hh通路的激活状态是普遍的,与病人的淋巴结转移无关,与癌肿的大小也无关。但与乳腺癌的特殊的病理类型可能有关。所有的64例肿瘤标本与周围正常组织相比都表现为Gli1强阳性染色,核染色Glil阳性/所有的肿瘤细胞的百分比从2%～95%不等,中位数为40.87%。Gli的核染色与雌激素受体阳性相关（P<0.05）,与孕激素受体阳性无关。结论：Hedgehog(Hh)信号传导通路可以作为潜在的乳腺癌治疗的新颖靶点。     

梓醇调控Hedgehog信号通路促进BMSCs向成骨分化的实验研究

目的观察梓醇对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向成骨细胞分化的影响,并从Hedgehog信号角度探讨其可能的作用机制。方法体外分离培养BMSCs,并通过流式细胞仪对细胞加以鉴定。pNPP法筛选梓醇最佳促BMSCs骨向分化的浓度,以最佳梓醇浓度干预BMSCs的成骨性分化。后续实验分4组:对照组、梓醇组、经典组和联合组(梓醇组+经典组)。酶标法检测碱性磷酸酶(ALP)活性,茜素红染色法检测矿化结节数目,QPCR法检测Shh mRNA、Ihh m RNA水平,Western blot法检测Ptch1、Smo及Gli1蛋白的表达。结果梓醇促BMSCs骨向分化的最佳浓度为1×10-4mol/L;与对照组比较,梓醇组的ALP活性显著增强,矿化结节数目显著升高,Shh mRNA水平显著上升,Ptch1、Smo及Gli1蛋白表达均显著升高(P0.05);与经典组比较,梓醇组的ALP活性、矿化结节数目、Shh m RNA水平、Ptch1、Smo和Gli1蛋白表达均低于经典组(P0.05);各组中Ihh mRNA水平无明显变化。结论梓醇能够促进BMSC向成骨分化,可能与上调Hedgehog信号传导通路相关分子有关。     

子宫颈癌中Hedgehog信号通路蛋白表达与人乳头状瘤病毒16型感染的关系

目的 探讨Hedgehog(Hh)信号通路蛋白在宫颈癌及其癌前病变中的临床病理学意义,并分析其与人乳头状瘤病毒(HPV)16型感染的关系.方法 32例正常宫颈上皮、71例宫颈上皮内瘤变(CIN;CIN Ⅰ 28例,CIN Ⅱ 18例,CIN Ⅲ25例)和80例宫颈鳞状细胞癌共183例选自延边大学医院、延边妇幼保健院和延边肿瘤医院病理科存档蜡块.应用PCR技术检测上述组织中HPV16型的感染情况,并应用Shh、Ihh、Ptch和Smo 4种Hh信号通路蛋白抗体、组织芯片和免疫组织化学EnVision法检测Hh信号通路蛋白在上述病变组织中的表达情况.结果 Shh、Ihh、Ptch和Smo在正常宫颈黏膜上皮中为弱阳性,而在宫颈癌和CIN Ⅲ中呈强阳性,其表达率均显著高于正常宫颈黏膜上皮(P均<0.05).80例官颈癌标本中HPV16阳性率是77.5%(62/80),而且Shh蛋白的强阳性率在HPV16型阳性的宫颈癌组织中显著高于HPV16阴性组(P<0.05).结论 Hh信号通路蛋白过表达可以作为宫颈癌及其癌前病变的早期辅助诊断指标并有望成为宫颈癌靶向治疗的新靶点,而且Shh蛋白的过表达与HPV16型感染密切相关.     

Sonic hedgehog信号通路在单侧输尿管梗阻大鼠肾组织中的表达变化及意义

 目的：探讨Sonic hedgehog（Shh）信号通路在单侧输尿管梗阻（UUO）大鼠肾组织中的表达变化及意义。方法：将48只SD大鼠随机分为UUO模型组（n=24）和假手术组（n=24）,梗阻术3、7和14 d后取其梗阻侧肾脏组织。用HE和Masson染色检测肾间质纤维化程度,免疫组织化学染色检测Shh通路分子Shh、Ptch1、Smo、Gli1及III型胶原的蛋白表达,酶联免疫吸附实验（ELISA）检测肾组织中TGF-β₁和Shh含量,real-time RT-PCR检测TGF-β₁、I和III型胶原及Shh通路分子mRNA表达。结果：HE和Masson染色显示,梗阻侧肾组织出现明显的纤维化病变,且随时间延长而加剧。TGF-β₁、I和III型胶原含量在梗阻肾中表达明显增高（P<0.05）。同时,Shh信号通路分子Shh、Smo和Gli mRNA和蛋白在梗阻肾中表达明显升高（P<0.05）,而Ptch1 mRNA和蛋白的表达下调（P<0.01）,提示Shh信号被激活。相关分析表明,Shh信号起始信号Shh水平的升高与TGF-β₁含量增加呈明显的相关。结论：UUO大鼠诱导肾间质纤维化发生过程中,Shh信号通路分子被激活,推测可能的机制是活化的Shh信号通路诱导TGF-β₁表达和释放,导致肾间质纤维化。     

干扰miR-31表达初期Notch和Hedgehog信号通路相关基因在神经干细胞中的表达变化

目的:研究干扰miR-31表达初期Notch和Hedgehog信号通路相关基因在神经干细胞(NSCs)中的表达变化。方法:利用荧光定量PCR对干扰miR-31表达初期Notch和Hedgehog信号通路相关基因在NSCs中的表达变化进行研究。结果:干扰与过表达miR-31后3 d,NSCs中的Notch信号通路相关基因Notch2的表达均增加,Jag2、Dll3和Hes1等的表达均降低;Hedgehog信号通路相关基因Wnt3的表达均增加,Bmp5与Wnt7a的表达均降低。结论:影响miR-31的表达可引发NSCs发生分化,在此过程中Notch与Hedgehog信号通路中几个基因的表达都产生相应改变,表明miR-31与NSCs分化过程相关。     

类风湿关节炎患者外周血单个核细胞Sonic Hedgehog信号通路表达的初步研究

目的: 初步探讨类风湿关节炎(RA)患者外周血单个核细胞(PBMCs)和滑膜组织中Sonic Hedgehog(Shh)信号通路相关因子表达及意义。方法: 收集符合1987年美国风湿病学会(ACR)RA分类标准、28个关节疾病活动度评分(DAS28)≥3.2,病情活动RA患者(35例)及年龄、性别相匹配的健康志愿者(35例)外周血2 mL,分离PBMCs,提取总RNA,采用实时荧光定量PCR(real-time PCR)检测Shh信号通路中信号肽Shh、膜受体Ptch1和核转录因子Gli1 mRNA的表达。收集10例病情中度活动(DAS28≥3.2)RA患者的滑膜组织,同时收集5例外伤或半月板损伤(无关节炎)者滑膜组织作为对照组,免疫组化检测Shh、Ptch1和Gli1的蛋白表达情况。结果: RA患者PBMCs中Shh和Gli1 mRNA的相对表达量分别为1.36±1.48和1.15±0.68,对照组上述信号分子的mRNA表达量分别为0.47±0.25和0.49±0.05,2组差异有统计学意义(P<0.05),Ptch1 mRNA表达在2组间的差异无统计学意义(P>0.05);免疫组化示Shh和Gli1蛋白表达的阳性细胞百分率均高于对照组(P<0.05),Ptch1蛋白表达阳性细胞百分率2组无显著差异(P>0.05)。结论: RA患者PBMCs与滑膜组织中检测到Shh通路相关信号分子Shh和Gli1的表达上调,提示RA患者中可能存在Shh信号通路的激活,其在RA发病机制中的作用值得进一步研究。     

SD大鼠肝细胞癌模型Hedgehog信号通路与复方苦参干预后的效应

背景：有研究表明在人类肝细胞癌中存在Hedgehog信号通路异常激活,但在动物肝细胞癌中Hedgehog信号通路是否激活尚不清楚。苦参可抑制肝癌细胞增殖周期,但其与Hedgehog信号通路的关系少见报道。 目的：观察Hedgehog信号通路在人工诱导SD大鼠肝细胞癌中的作用,以及复方苦参药物对Hedgehog信号通路影响。 方法：二乙基亚硝胺诱导SD大鼠建立肝细胞癌大鼠模型后,随机分为模型组、低剂量苦参组和高剂量苦参组,并设立空白对照组。建模后15周,分别在低、高剂量苦参组腹腔注射30,90 mg/(kg•d)复方苦参治疗3周后,应用免疫组织化学染色和RT-PCR法检测这所有大鼠组织中在Hedgehog信号通路中靶基因转录因子Gli2和跨膜蛋白受体复合物Ptch的表达情况。 结果与结论：相比对照组,模型组、低剂量和高剂量苦参组肝癌组织中Gli2和Ptch均呈高表达(P < 0.01)。结果提示,在人工诱导的SD大鼠肝癌模型中,Hedgehog信号通路处于激活状态。在给予高低剂量复方苦参干预下,Hedgehog信号通路靶基因转录因子Gli2和跨膜蛋白受体复合物Ptch呈高表达。     

Surgical outcome of synchronous second primary cancer in patients with gastric cancer

PURPOSE: In order to improve the likelihood of curative and safe gastric surgery, this study investigated the clinical features and surgical outcomes of gastric cancer with a synchronous cancer. PATIENTS AND METHODS: The clinicopathological data of 10,090 gastric cancer patients at Samsung Medical Center from September 1994 to December 2006 were retrospectively analyzed. Of them, 90 patients with gastric cancer and a synchronous second primary cancer underwent simultaneous surgery for gastric cancer and second primary cancer. The clinicopathological characteristics of the patients, surgical outcome, and prognosis were examined. RESULTS: The most common synchronous second primary cancer was colorectal cancer (37 patients), followed by hepatocellular carcinoma (13 patients), renal cell carcinoma (11 patients), and pancreatic carcinoma (5 patients). The incidence of a second primary cancer in the gastric cancer patients was higher than the incidence in the general population. Stage I gastric cancer patients had more synchronous cancers than stage II patients (59 vs. 31). Postoperative complications were encountered in 7 patients. Four patients underwent reoperation. Two patients died from hepatic failure and leakage of esophagojejunal anastomosis. The 5-year survival rate of stage I and II gastric cancer was 61% and 39%, respectively. CONCLUSION: Since gastric cancer patients with a synchronous second primary cancer are not rare, the possibility of synchronous cancers in gastric cancer patients should be considered. The prognosis of early stage gastric cancer patients with a synchronous second primary cancer was influenced more by the presence of the second primary cancer than by the gastric cancer itself.     

Ternary cancer comprising prostatic cancer,esophageal cancer,and gastric cardia cancer: a case report and literature review

A 61-year-old male patient was admitted to our hospital with frequent urination, urgency, and increased nocturia for more than 3 months, and the symptoms were aggravated for 1 week. Prostate biopsy revealed prostatic adenocarcinoma. After 5 months, the patient developed dysphagia and gastroscopy showed a middle and lower esophageal cancer (squamous cell carcinoma). 12 months later, he returned to the hospital because of dysphagia. He was examined by gastroscopy which showed the cardia to have low-grade adenocarcinoma. The patient was given Casodex + Zoladex endocrine therapy, zoledronic acid inhibiting bone destruction, concurrent chemoradiotherapy, capecitabinen tablets at a dose of 1000 mg bid, 3 cycles of intravenous paclitaxel at 180 mg/d1 plus cisplatin 60 mg/d1-2, 4 cycles of intravenous paclitaxel at 150 mg/d1 plus cisplatin at 60 mg/d1 as systemic chemotherapy. The curative effect is was considerable after treatment, and the patient’s condition was stable. Since the onset of the disease in March 2018, the patient’s condition had not progressed significantly for 27 months. The diagnosis and treatment of this patient with ternary cancer in the hospital improved the clinician’s understanding of multiple primary cancers. Multidisciplinary treatment improved the patient’s prognosis and quality of life. We reviewed similar case reports and retrospective studies of multiple primary cancers and found that there is no specific treatment for multiple primary cancers, but a corresponding treatment program can be formulated for each tumor to control progression while screening for possible other primary tumors.     

间质干细胞、肿瘤干细胞和肿瘤

近来研究认为肿瘤来源于干细胞,提出了肿瘤中存在极少量肿瘤干细胞(CSC)的新学说.问质干细胞(MSC)作为间质细胞的来源,与肿瘤的关系尤为密切.本文就间质干细胞,肿瘤干细胞和肿瘤的发生,发展作一综述.     

Routine testing for PALB2 mutations in familial pancreatic cancer families and breast cancer families with pancreatic cancer is not indicated

PALB2-mutation carriers not only have an increased risk for breast cancer (BC) but also for pancreatic cancer (PC). Thus far, PALB2 mutations have been mainly found in PC patients from families affected by both PC and BC. As it is well known that the prevalence of gene mutations varies between different populations, we studied the prevalence of PALB2 mutations in a Dutch cohort of non-BRCA1/2 familial PC (FPC) families and in non-BRCA1/2 familial BC (FBC) families with at least one PC case. Mutation analysis included direct sequencing and multiplex ligation-dependent probe amplification (MLPA) and was performed in a total of 64 patients from 56 distinct families (28 FPC families, 28 FBC families). In total, 31 patients (48%) originated from FPC families; 24 were FPC patients (77%), 6 had a personal history of BC (19%) and 1 was a suspected carrier (3.2%). The remaining 33 patients (52%) were all female BC patients of whom 31 (94%) had a family history of PC and 2 (6.1%) had a personal history of PC. In none of these 64 patients a PALB2 mutation was found. Therefore, PALB2 does not have a major causal role in familial clustering of PC and BC in non-BRCA1/2 families in the Dutch population.     

Special cancer microenvironment in human colonic cancer: Concept of cancer microenvironment formed by peritoneal invasion (CMPI) and implication of subperitoneal fibroblast in cancer progression

Clinical outcomes of colorectal cancer are influenced not by tumor size, but by spread into the bowel wall. Although assessment of serosal involvement is an important pathological feature for classification of colon cancer, its diagnostic consistency has been questioned. Using elastic staining, we assessed elastic laminal invasion (ELI) for more objective stratification of deep tumor invasion around the peritoneal surface. In addition, pathological characteristic features of marked tumor budding, fibrosis, and macrophage infiltration in the tumor area with ELI was elucidated. This characteristic tumor area was termed cancer microenvironment formed by peritoneal elastic laminal invasion (CMPI). We elucidated histoanatomical layer‐dependent heterogeneity of fibroblast in colonic tissue. Furthermore, subperitoneal fibroblasts (SPFs) play a crucial role in tumor progression and metastasis in CMPI. Our ELI and CMPI concept contributes not only to objective pathological diagnosis, but also sheds light on biological research of special cancer microenvironments detectable in human colorectal cancers. Herein, we describe the diagnostic utility of ELI and morphological alteration in advanced colorectal cancers to determine the phenomenon that occurs when tumors invade around the peritoneal surface. Next, biological research of CMPI is reviewed to stress the importance of pathological research to establish new biological concepts.     

The risk of contralateral breast cancer in daughters of women with and without breast cancer

We aimed to estimate the 15‐year and lifetime risks of contralateral breast cancer in breast cancer patients according to the age of diagnosis of the first cancer and the history of breast cancer in the mother. The risks of contralateral breast cancer were estimated for all 78,775 breast cancer patients in the Swedish Family‐Cancer Database (age at diagnosis of first breast cancer <70 years). The risk of experiencing a contralateral breast cancer within 15 years of diagnosis was 8.4% [95% confidence interval (CI): 8.1–8.7%] for women with an unaffected mother, was 12% (95%CI: 11–13%) for a woman with a mother with unilateral breast cancer and was 13% (95%CI: 9.5–17%) for women with a mother with bilateral breast cancer. In early‐onset diagnosed women (<50 years) with an unaffected mother, the risk of contralateral breast cancer until age 80 was 23% (95%CI: 20–26%) and for late‐onset (50–69 years) diagnosed women it was 17% (95%CI: 14–21%). In a woman with a mother with an early‐onset unilateral breast cancer, risk of contralateral breast cancer by age 80 was 35% (95%CI: 25–46%). Women with a mother with early‐onset bilateral breast cancer had 31% (95%CI: 12–67%) lifetime risk of contralateral breast cancer. The risk of contralateral breast cancer is higher for daughters of breast cancer patients than for daughters of women without breast cancer. Maternal cancer history and age at onset of first breast cancer in women should be taken into account when counseling breast cancer patients about their risk of contralateral breast cancer.     

Diagnosis analysis in breast cancer and cervical cancer screening

Assessment of the accuracy of diagnostic procedures has been made independent of the diagnostic criteria used by means of Relative Operating Characteristics (ROC) analysis. A ROC curve describes the mutual relationship between the sensitivity and specificity of a diagnostic decision on the basis of various diagnostic criteria. The construction of such ROC curves is made possible if diagnoses are graded into levels of certainty. The curve enables the choice of an operating point with predetermined sensitivity and specificity values for the diagnosis decision. The population-based breast-cancer and cervical cancer screening projects carried out in Utrecht demonstrated an excellent fit between actual data and the calculated ROC curves. Analysis of the accuracy or performance of cytological diagnosis uncovered a problem arising from the similarly graded histopathological reference criteria used to determine the 'truth' of the cytological diagnosis decisions. The proposed solution is a serial calculation of ROC curves, one for each level differentiating between the histopathological categories. The ensuing three-dimensional ROC hill may reveal a summit marking numerically advantageous diagnosis criterion levels for both the test and the disease to be detected, or a depression signalling locally below-standard detection performance.     

肿瘤干细胞的研究现状

传统理论认为肿瘤生长是所有肿瘤细胞一起增殖的结果,因而治疗方法主要是针对肿瘤组织内的大多数细胞,常有复发和转移,使治疗失败。肿瘤干细胞的提出,可靶向性杀伤肿瘤干细胞从而使根治肿瘤和防止肿瘤复发和转移成为可能,且先前诸多关于肿瘤发生发展机制、细胞信号途径等研究成果可能需要重新评价,对传统的肿瘤治疗方式提出了巨大挑战。就肿瘤干细胞的起源、存在证据、肿瘤干细胞与正常干细胞之间关系、临床意义及展望综述如下。