Single-cell transcriptomics reveals heterogeneous progression and EGFR activation in pancreatic adenosquamous carcinoma

Pancreatic adenosquamous carcinoma (PASC) — a rare pathological pancreatic cancer (PC) type — has a poor prognosis due to high malignancy. To examine the heterogeneity of PASC, we performed single-cell RNA sequencing (scRNA-seq) profiling with sample tissues from a healthy donor pancreas, an intraductal papillary mucinous neoplasm, and a patient with PASC. Of 9,887 individual cells, ten cell subpopulations were identified, including myeloid, immune, ductal, fibroblast, acinar, stellate, endothelial, and cancer cells. Cancer cells were divided into five clusters. Notably, cluster 1 exhibited stem-like phenotypes expressing UBE2C, ASPM, and TOP2A. We found that S100A2 is a potential biomarker for cancer cells. LGALS1, NPM1, RACK1, and PERP were upregulated from ductal to cancer cells. Furthermore, the copy number variations in ductal and cancer cells were greater than in the reference cells. The expression of EREG, FCGR2A, CCL4L2, and CTSC increased in myeloid cells from the normal pancreas to PASC. The gene sets expressed by cancer-associated fibroblasts were enriched in the immunosuppressive pathways. We demonstrate that EGFR-associated ligand-receptor pairs are activated in ductal-stromal cell communications. Hence, this study revealed the heterogeneous variations of ductal and stromal cells, defined cancer-associated signaling pathways, and deciphered intercellular interactions following PASC progression.     

A case of maxillary sarcoma in a chimpanzee (Pan troglodytes)

Oral malignancy is rare in chimpanzees. A 34‐year‐old female chimpanzee (Pan troglodytes) at Kumamoto Sanctuary, Japan, had developed it. Treatment is technically difficult for chimpanzees while malignant neoplasm is seemingly rising in captive populations. Widespread expert discussion, guidelines for treatment, especially for great apes in terminal stages is urgently needed.     

Targeting chemokine pathways in esophageal adenocarcinoma

Esophageal adenocarcinoma (EAC) is one of the fastest growing malignancies in the US and needs newer therapeutic and diagnostic strategies. Chronic inflammation plays a role in the pathogenesis of EAC and contributes to the dysplastic conversion of normal esophageal epithelium to Barrett's esophagus and frank adenocarcinoma. Chemokines play important roles in mediating inflammation and recent evidence implicates these ligands and their receptors in the development and spread of various tumors. We demonstrated that the chemokines IL8, CXCL1 and CXCL3 are significantly overexpressed during esophageal carcinogenesis and accompanied by amplification and demethylation of the chr4q21 gene locus. We also demonstrated that IL8 levels can be detected in serum of patients with EAC and can serve as potential biomarkers. We now demonstrate that inhibition of IL8 receptor, CXCR2, leads to decreased invasiveness of esophageal adenocarcinoma derived cells without affecting cellular proliferation. Taken together, these studies reveal the important roles that chemokines play in development of esophageal cancer and demonstrate that these pathways can serve as potential therapeutic targets.     

Ezrin在胃癌组织中的表达及与Hp-L型感染关系的探讨

目的通过检测胃癌组织中幽门螺杆菌L(Helicobacter pyloriL-form,Hp-L)型感染以及Ezrin的表达情况,探讨Hp-L型、Ezrin在胃癌组织中的表达及临床意义。方法 (1)应用革兰染色法和免疫组织化学Elivision法检测80例胃癌组织和40例对照组织中的Hp-L型感染情况。(2)应用免疫组织化学Elivision法检测上述各组织中Ezrin蛋白的表达。(3)应用逆转录多聚酶链反应(RT-PCR)法检测30例新鲜胃癌组织及与其相对应的30例远端切缘正常组织中Ezrin mRNA的表达。结果 (1)胃癌组中革兰染色L型的检出率为80.00%(64/80)、免疫组化Hp-L型阳性率81.25%(65/80),两种方法检测的结果具有一致性(P0.05)。80例胃癌组织中Hp-L型阳性(即革兰染色L型检出阳性和免疫组化Hp-L型抗原表达同时阳性)的病例数为56例,其阳性率为70.00%;对照组中革兰染色L型检出率为22.50%(9/40),免疫组化Hp-L型阳性率40%(16/40)二者检测结果也具有一致性(P0.05)。40例对照组织中Hp-L型阳性例数为9例,其阳性率为22.50%。胃癌组与对照组的Hp-L型阳性率相比,差异具有统计学意义(P0.05);(2)胃癌组Hp-L型感染阳性率仅与胃癌的淋巴结转移有关(P0.05),而与其他临床病理因素无关;(3)RT-PCR法和免疫组织化学Elivision法显示胃癌组中Ezrin mRNA及Ezrin蛋白的表达均高于对照组(P0.05),且经统计学分析发现Ezrin表达水平与胃癌细胞的分化程度、浸润深度及淋巴结转移有关(P0.05),而与临床分期、患者的年龄及性别无关(P0.05);(4)胃癌中Hp-L型阳性组的Ezrin蛋白表达阳性率71.43%(40/56)高于Hp-L阴性组54.17%(13/24)(P0.05),且Hp-L型阳性和Ezrin蛋白阳性呈正相关(r=0.456,P0.05)。结论 Hp-L型感染阳性率和Ezrin表达阳性率在胃癌中均较高,二者可能协同促进胃癌的发生发展及浸润转移。     

PTEN在人食管上皮永生化细胞株和食管癌细胞株中的表达

目的:探讨人胚食管上皮永生化细胞株SHEE、SHEEMT、食管癌细胞株EC8712中PTEN表达的差,判断食管上皮细胞在恶性转化过程中是否有PTEN缺失现象的发生;方法:培养三种细胞株,采用免疫组织化学、激光共聚焦显微镜、流式细胞仪和Westernblot等检测方法对三种细胞株中PTEN的表达进行检测。结果:三种细胞株均有PTEN的表达,表达强度与分化程度有关,PTEN在三种细胞株中表达强弱顺序为SHEE>SHEEMT>EC8712,差异有统计学意义(P<0.01);结论:PTEN在SHEE、SHEEMT和EC8712三种来源于食管鳞状上皮但分化程度不同的细胞株均中表达,表达强度与分化程度相关,分化程度越高,表达越强。     

再生基因家族的研究

再生基因家族自1988年被发现以来,其在糖尿病、炎症创伤与肿瘤尤其在消化系统肿瘤中的作用日渐被重视。越来越多的该家族成员被发现,并已开始考虑在临床治疗中应用。这些研究开始显示再生基因家族的潜在应用价值。 Abstract:Since the first member of Reg gene was discovered in 1988,it has been verified that Reg genes play important roles in diabetes,inflammation and injury,and tumors.More members were cloned and their application in treatment was studied.With the development of related research,there is a great potential of Reg family in biomedical field.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Characterization of Glycoproteoforms of Integrins α2 and β1 in Megakaryocytes in the Occurrence of JAK2V617F Mutation-Induced Primary Myelofibrosis

Primary myelofibrosis (PMF) is a neoplasm prone to leukemic transformation, for which limited treatment is available. Among individuals diagnosed with PMF, the most prevalent mutation is the JAK2V617F somatic point mutation that activates the Janus kinase 2 (JAK2) enzyme. Our earlier reports on hyperactivity of β1 integrin and enhanced adhesion activity of the α2β1 complex in JAK2V617F megakaryocytes (MKs) led us to examine the new hypothesis that this mutation leads to posttranslational modification via changes in glycosylation. Samples were derived from immunoprecipitation of MKs obtained from Vav1-hJAK2^V617F and WT mice. Immunoprecipitated fractions were separated by SDS-PAGE and analyzed using LC-MS/MS techniques in a bottom-up glycoproteomics workflow. In the immunoprecipitate, glycopeptiforms corresponding to 11 out of the 12 potential N-glycosylation sites of integrin β1 and to all nine potential glycosylation sites of integrin α2 were observed. Glycopeptiforms were compared across WT and JAK2V617F phenotypes for both integrins. The overall trend observed is that JAK2V617F mutation in PMF MKs leads to changes in β1 glycosylation; in most cases, it results in an increase in the integrated area of glycopeptiforms. We also observed that in mutated MKs, changes in integrin α2 glycosylation were more substantial than those observed for integrin β1 glycosylation, a finding that suggests that altered integrin α2 glycosylation may also affect activation. Additionally, the identification of proteins associated to the cytoskeleton that were co-immunoprecipitated with integrins α2 and β1 demonstrated the potential of the methodology employed in this study to provide some insight, at the peptide level, into the consequences of integrin activation in MKs. The extensive and detailed glycosylation patterns we uncovered provide a basis for future functional studies of each site in control cells as compared to JAK2V617F-mutated cells. Data are available via ProteomeXchange with identifier PXD030550.     

Gynaecological cancers and their cell lines

Cell lines are widely used for various research purposes including cancer and drug research. Recently, there have been studies that pointed to discrepancies in the literature and usage of cell lines. That is why we have prepared a comprehensive overview of the most common gynaecological cancer cell lines, their literature, a list of currently available cell lines, and new findings compared with the original studies. A literature review was conducted via MEDLINE, PubMed and ScienceDirect for reviews in the last 5 years to identify research and other studies related to gynaecological cancer cell lines. We present an overview of the current literature with reference to the original studies and pointed to certain inconsistencies in the literature. The adherence to culturing rulesets and the international guidelines helps in minimizing replication failure between institutions. Evidence from the latest research suggests that despite certain drawbacks, variations of cancer cell lines can also be useful in regard to a more diverse genomic landscape.     

Esophageal cancer lymph node metastasis–associated gene signature optimizes overall survival prediction of esophageal cancer

Esophageal cancer (EC) is characteristic of early regional lymph node metastasis (LNM) and most patients with metastasis have a poor prognosis. However, the current diagnostic techniques do not enable precise differentiation of EC LNM, prognostic stratification, and individual survival estimation. To identify potential molecular biomarkers for EC patients with LNM, we explored differently expressed genes in The Cancer Genome Atlas database between 77 non-LNM cases and 88 LNM cases by limma package R. Then, according to univariate and multivariate Cox regression analyses, we constructed an 8-messenger RNA (mRNA) prognostic signature model, which could predict the outcome in a more exact way. The area under the curve of the risk score is significantly higher than other clinical information, indicating that the 8-mRNA–based risk score is a good indicator for prognosis. Then, combined with other individual risk factors, such as age, sex, T stage, M stage, etc, we could precisely calculate the individual 1-, 3-, and 5-year survival rates. The Gene Set Enrichment Analysis, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes analysis indicate that the risk model is mainly associated with cancer-related pathways, such as cell division, cellular meiosis, and cell cycle regulation. In summary, the 8-mRNA–based risk score model that we developed successfully predicts the survival of EC. It is independent of clinical information and performing better than other clinical information for prognosis.