Uptake of bacterial lipopolysaccharide and expression of tumor necrosis factor α mRNA in isolated rat intrahepatic bile duct epithelial cells *

AIM To study the uptake of bacterial lipopolysaccharides (LPS) and expression of tumor necrosis factor α mRNA (TNF α mRNA) with cultured rat intrahepatic bile duct epithelial cells.

Relationship between the GH-IGFs axis and the proliferation of bile duct cancer cell line QBC939 in vitro

BACKGROUND: In recent years, recombined human growth hormone (rhGH) has been increasingly used in patients to help them recover from operation. But GH, as a mitogen, can promote cell renewal and increase malignant transformation. In the current study, we assessed the proliferation of a bile duct cancer cell line (QBC939) in vitro with GH and explored the possible relationship with the axis of GH-IGFs (insulin-like growth factors). METHODS: QBC939 cells in the exponential growth stage were harvested and divided into an experimental group (GH group) and a control group (NS group). The GH group was divided into four sub-groups according to the dose of GH and culture time (50 mu g/L for 2 hours, 50 mu g/L for 24 hours, 100 mu g/L for 2 hours, 100 mu g/L for 24 hours). The NS group was divided into two sub-groups (NS for 2 hours and NS for 24 hours). After 2 or 24 hours, IGF-1 and IGF-2 were detected using the enzyme-linked immunosorbent assay. The QBC939 cells cultured for 24 hours with two GH concentrations were made into single cell suspensions and samples underwent subsequent cell cycle evaluation. Messenger RNA of IGF-1 and IGF-2 receptor (IGF-1RmRNA and IGF-2RmRNA) were tested with the method of in situ hybridization. RESULTS: There was no statistically significant difference between the GH and NS groups after 2 hours of culture (P>0.05). But after 24 hours of culture, GH stimulated cell growth in vitro and also elevated the percentage in S phase and the proliferation index (P<0.05). IGF-1RmRNA and IGF-2RmRNA were expressed in QBC939 in contrast to the blank group. The expression of IGF-1RmRNA increased with the dose of GH, but IGF-2RmRNA did not. CONCLUSION: GH can stimulate QBC939 cell growth and proliferation in vitro and the mechanism is most likely by the GH-IGF-1-IGF-1R axis.

Benign giant-cell tumor of the common bile duct:A case report

Primary giant-cell tumors rarely arise in the common bile duct. We herein report a case of primary giant-cell tumor of the common bile duct. The patient was an 81-year-old male who was diagnosed with a well-defined 1.2-cm mass projecting into the lumen of the middle common bile duct. Excision of the gallbladder and extrahepatic bile duct and a Roux-en-Y cholangiojejunostomy were performed. Histologically, the tumor had no association with carcinomas of epithelial origin and was similar to giant-cell tumors of the bone. The tumor consisted of a mixture of mononuclear and multinucleated osteoclast-like giant cells. The mononuclear cells showed no atypical features, and their nuclei were similar to those of the multinucleated giant cells. CD68 was expressed on the mononuclear and multinucleated osteoclast-like giant cells, whereas CD163 immunoreactivity was restricted to the mononuclear cells. Six months after the operation, the patient was still alive and had no recurrence. The interest of this case lies in the rarity of this entity, the difficulty of preoperative diagnosis, and this tumor&#x02019;s possible confusion with other malignant tumors.

The abnormal expression of E-cadherin in intrahepatic bile duct epithelia cells in biliary atresia and its relationship with apoptosis

To explore the relationship between the expression of E-cadherin and the apoptosis in intrahepatic bile duct epithelial cells in biliary atresia (BA).Methods The E-cadherin expression was demonstrated by immunohistochemical staining for the liver specimens from 38 children with BA and 16 normal children.The apoptotic intrahepatic bile duct epithelial cells in these specimens were visualized by TdT-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay,and the apoptotic index (AI) was calculated from the percentage of apoptotic cells in total cells.Results The intensity of E-cadherin expression in bile duct epithelial cells in BA group was lower than that in the normal control group (0.33±0.12 vs 0.62±0.20,P<0.01).On the other hand,the AI in BA group was significant higher than that in control group (51.74±19.93 vs 12.34±19.32,P<0.01).An inverse correlation was detected between the intensity of E-cadherin and the AI in the liver from children with BA.Conclusion The abnormal decrease of E-cadherin may lead to an increase of the apoptosis of intrahepatic bile epithelial cells in BA,resulting in developmental disorder of intrahepatic bile duct and ductal plate malformation in the liver.12 refs,4 figs,1 tab.

Neuroendocrine carcinoma of the extrahepatic bile duct: Case report and literature review

Neuroendocrine carcinoma (NEC) of the extrahepatic bile duct is rare, and only 22 cases have been reported. Only two of these were large-cell NEC (LCNEC); the vast majority were small-cell NEC. Here, we report a third case of LCNEC of the extrahepatic bile duct. A 76-year-old male presented to a local hospital with painless jaundice. Imaging studies revealed a tumor at the hepatic hilum. The patient underwent right hepatic lobectomy, bile duct resection, and cholecystectomy. The resection specimen showed a 5.0-cm invasive neoplasm involving the hilar bile ducts and surrounding soft tissue. Histologically, the tumor consisted of nests of medium to large cells with little intervening stroma. The tumor invaded a large portal vein branch. All four excised lymph nodes were positive for metastasis, and metastatic deposits were also present in the gallbladder wall. The tumor was diffusely positive for synaptophysin and focally positive for chromogranin A. Approximately 70%-80% of the tumor cells were positive for Ki-67, indicating strong proliferative activity. A diagnosis of LCNEC was made. A few bile ducts within and adjacent to the invasive tumor showed dysplasia of the intestinal phenotype and were focally positive for synaptophysin and chromogranin A, suggesting that the dysplastic intestinal-type epithelium played a precursor role in this case. A postoperative computer tomography scan revealed rapid enlargement of the abdominal and retroperitoneal lymph nodes. The patient died 21 d after the operation. NEC of the bile duct is an aggressive neoplasm, and its biological characteristics remain to be better defined.

Effects of LPS-induced cholangitis on the cytoskeleton morphology of bile duct epithelium and the intervention mechanism of Dahuang Lingxian formula for these changes

Objective:To observe the effect of lipopolysaccharide(LPS)on cytoskeleton and the effect of Dahuang Lingxianfang on nF-KB/MAPK signaling pathway.Methods:Biliary epithelial cells of each group were stained with photolipin fluorescent staining,and the arrangement of cytoskeleton was observed under laser confocal microscope.Western blotting was used to detect the expression of f-actin.Results:After LPS intervention,the biliary epithelial cells showed nuclear shrinkage or damage,and the skeleton was broken or lumped.The cytoskeleton was partially repaired after the intervention of pathway blocking preparation combined with RHUbarb Lingxianfang.All the other groups had different degree of cytoskeleton fracture.Compared with normal group,the expression of F-actin protein in LPS group was decreased(P<0.05);Compared with LPS group,the expression of F-actin in LPS+TCM group,LPS+PDTC+TCM group,LPS+SB203580+TCM group and LPS+PDTC+SB203580+TCM group was significantly increased(P<0.05);Compared with traditional Chinese medicine group,the expression of F-actin in LPS+PDTC+traditional Chinese medicine group,LPS+SB203580+traditional Chinese medicine group and LPS+PDTC+SB203580+traditional Chinese medicine group had no significant difference(P>0.05).Conclusion:RHUbarb Lingxianfang can restore the sequence of biliary epithelial cytoskeleton and protect its microfilament structure under inflammation,and its mechanism may be related to the regulation of NF-KB/MAPK signaling pathway.

Apoptosis and proliferation of intrahepatic bil educt after ischemia-reperfusion injury

BACKGROUND: In orthotopic liver transplantation, ische-mic-reperfusion is one of the most important factors thatcause the incidence of biliary complicance. The aim of thisstudy was to investigate the effects of ischemia reperfusionon epithelial cells apoptosis and proliferation of intrahepaticbile duct (IBD) (>20 μm).METHODS: 30-minute warm ischemia was applied to ratlivers respectively, and experiment was performed on days2,7, 14, 28 after reperfusion. Apoptosis was determined insitu by morphology and TUNEL, and cholangiocyte proli-feration was evaluated in situ by morphometry of liver sec-tions stained for cytokeratin-19 ( CK-19) and by prolifera-ting cellular nuclear antigen staining in liver sections.RESULTS: Two days after ischemia reperfusion, apoptosisof cells was observed in large intrahepatic bile ducts (>20μm) (5.6%±1.2%) , but the number of large intrahepaticbile ducts reduced (0.32 ±0.06). Seven days after ischemiareperfusion, the apoptosis index of cholangiocytes de-creased to 1.2%±0.3%, and the number of intrahepatic bileducts began to proliferate and returned to nearly normal onday 28.CONCLUSION: Ischemia reperfusion causes a decrease inthe number of intrahepatic bile ducts (>20 μm) as a resultof a higher rate of apoptosis and absence of initial prolifera-tion.

Raddeanin A promotes apoptosis and ameliorates 5-fluorouracil resistance in cholangiocarcinoma cells

BACKGROUND Bile duct cancer is characterized by fast metastasis and invasion and has been regarded as one of the most aggressive tumors due to the absence of effective diagnosis at an early stage.Therefore,it is in the urgent demand to explore novel diagnostic approaches and therapeutic strategies for bile duct cancer to improve patient survival.Raddeanin A(RA)is extracted from the anemone raddeana regel and has been demonstrated to play antitumor roles in various cancers.AIM To investigate the effects of RA treatment on bile duct cancer cells.METHODS In this study,four cholangiocarcinoma cell lines(RBE,LIPF155C,LIPF178C,and LICCF)treated with RA were used to test the cell viability.The RA-associated cell functional analysis,5-fluorouracil(5-Fu)effectiveness as well as cell cycle-and apoptosis-related protein expression were investigated.RESULTS RA reduced cell viability in a dose-dependent pattern in four cell lines,and the migration and colony formation abilities were also impaired by RA in RBE and LIPF155C cell lines.RA sensitized cell lines to 5-Fu treatment and enhanced the effects of 5-Fu in cholangiocarcinoma.Also,RA decreased protein expression of Wee1,while the combinational effect of RA and 5-Fu decreased protein expressions of cyclooxygenase-2,B cell lymphoma 2,and Wee1 but increased protein levels of Bax,cyclin D1,and cyclin E.CONCLUSION Taken together,the results suggest that RA acts as an anti-cancer agent and enhancer of 5-Fu in bile duct cancer cells via regulating multiple cell cycle and apoptosis-related proteins.This finding provides novel clues to exploring a novel antitumor drug for bile duct cancer.

Small cell carcinoma of the liver and biliary tract without jaundice

An 80-year-old woman presenting with chest pain was found to have a large,lobulated soft tissue mass in the liver and nearby tissues on abdominal computed tomography(CT).The tumor had invaded the common hepatic artery and main portal vein.Jaundice developed 4 wk later,at which point,a pancreas and biliary CT scan revealed a large mass in the right lobe of the liver and a hilar duct obstruction,which was found to be a small cell carcinoma.Despite its rarity,liver and bile duct small cell carcinoma should be considered in the differential diagnosis of atypical chest pain without jaundice.

TTF-1 positive small cell cancers:Don't think they're always primary pulmonary!

Thyroid transcription factor 1 (TTF-1) plays a key role in morphogenesis of the lungs and is expressed in up to 90% of pulmonary small cell carcinomas.This explains why this marker is frequently used in the search for the primary origin of metastatic endocrine tumours.Here we report on a TTF-1 expressing mixed endocrine-exocrine carcinoma of the common bile duct in a patient with pulmonary nodules that did not appear to be neoplastic.TTF-1 positivity in pulmonary and extrapulmonary neuroendocrine tumours is reviewed,and we conclude that TTF-1 expression in neuroendocrine tumours of the small-cell type are not uncommon at extrapulmonary locations.Therefore,immunohistochem-istry for TTF-1 in such tumours should be interpreted with caution.

Preventive effects of autologous bone marrow mononuclear cell implantation on intrahepatic ischemic-type biliary lesion in rabbits

BACKGROUND: The ischemic-type biliary lesion (ITBL) is one of the most serious biliary complications of liver transplantation. This study aimed to investigate the effects of autologous bone marrow mononuclear cell (BM-MNC) implantation on neovascularization and the prevention of intrahepatic ITBL in a rabbit model. METHODS: The rabbits were divided into control, experimental model, and cell implantation groups, with 10 in each group. The model of intrahepatic ITBL was established by clamping the hepatic artery and common bile duct. Autologous BM-MNCs were isolated from the tibial plateau by density gradient centrifugation and were implanted through the common hepatic artery. Changes in such biochemical markers as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyltranspeptidase, total bilirubin and direct bilirubin were measured. Four weeks after operation, cholangiography, histopathological manifestations, differentiation of BM-MNCs, microvessel density and the expression of vascular endothelial growth factor were assessed. RESULTS: Compared with the experimental model group, the BM-MNC implantation group showed superiority in the time to recover normal biochemistry. The microvessel density and vascular endothelial growth factor expression of the implantation group were significantly higher than those of the control and experimental model groups. The ITBL in the experimental model group was more severe than that in the implantation group and fewer new capillary blood vessels occurred around it. CONCLUSIONS: Implanted autologous BM-MNCs can differentiate into vascular endothelial cells, promote neovascularization and improve the blood supply to the ischemic bile duct, and this provides a new way to diminish or prevent intrahepatic ITBL after liver transplantation. (Hepatobiliary Pancreat Dis Int 2010; 9:593-599)

基于关键免疫细胞亚群的胆管癌患者外周血早期诊断标志物的研究

目的基于关键免疫细胞亚群探究胆管癌患者外周血早期诊断标志物。方法于2021年5月至2023年5月选取在本院确诊为CCA的患者75例即为CCA组,同期在本院收治的75例胆管炎患者即为对照组。同期在本院体检健康的志愿者75例为健康组。采用流式细胞仪检测CD3^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+)水平;采用酶联免疫吸附试验(ELISA)法检测肿瘤坏死因子(TNF-α)、白细胞介素-2(IL-2)、γ干扰素(IFN-γ)水平。采用Pearson法分析CCA患者CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平与TNF-α、IL-2、IFN-γ水平的相关性;ROC曲线分析外周血免疫细胞亚群对CCA发生的诊断价值。多因素Logistic回归分析CCA发生的影响因素。结果与健康组相比,对照组、CCA组患者CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平显著升高;与对照组相比,CCA组患者CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平显著升高(P<0.05)。与健康组相比,对照组、CCA组患者IL-2、IFN-γ水平显著降低,TNF-α水平显著升高;与对照组相比,CCA组患者IL-2、IFN-γ水平显著降低,TNF-α水平显著升高(P<0.05)。相关性分析显示,CCA患者CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平与IL-2、IFN-γ呈负相关关系,与TNF-α呈正相关关系(P<0.05)。外周血免疫细胞亚群(CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)、TNF-α、IL-2、IFN-γ)联合检测对CCA发生诊断的AUC显著高于单一指标诊断的AUC的值(Z_(CD3^(+)vs联合)=4.424,P<0.001;Z_(CD4^(+)vs联合)=3.425,P=0.001;Z_(CD4^(+)/CD8^(+)vs联合)=4.502,P<0.001;Z_(TNF-αvs联合)=3.322,P<0.001;Z_(IL-2^(+)vs联合)=7.473,P=0.001;Z_(IFN-γ^(+)vs联合)=3.166,P=0.001)。多因素Logistic回归分析显示,CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)、IL-2、IFN-γ水平是影响CCA发生的影响因素(P<0.05)。结论CCA患者外周血免疫细胞亚群水平显著升高,联合检测能够提高对CCA发生的诊断效能。

Effects of substance P on growth of fibroblast-like cells derived from bile duct： an in vitro cell culture study

Background The possible role of substance P （SP） during wound healing has been the primary research focus in recent years,but its effect on the healing process after bile duct injury is little understood.This study aimed to investigate the effects of SP on growth of fibroblast-like cells derived from rabbit bile duct.Methods Fibroblast-like cells derived from rabbit bile duct were identified and divided randomly into control and experimental groups.SP-treated cells at different concentrations of 10^-9-10^-5 mol/L and control group were incubated,respectively,for 48 hours.After incubating,the effects of SP on cell proliferation were assessed by cell counts and MTT test.Apoptosis rate （AR） of cells was measured by flow cytometry.Results Cultured rabbit bile duct cells were fibroblast-like in morphology,and these cells were stained positively for vimentin and negatively for desmin.After SP was added to nonconfluent cells for 48 hours,cell numbers were significantly increased in experimental groups than in controls （P 〈0.05）.The maximum stimulation of cell proliferation was achieved at SP of 10^-5 mol/L.Bile duct fibroblast-like cells in the SP group showed a higher proliferating activity and lower AR than those in the control group or in the SP ＋ Spantide group （P 〈0.05）.Spantide partly inhibited the effects of SP on fibroblastlike cells.Examination under transmission electron microscopy revealed rough endoplasmic reticulum and prominent Golgi complexes after SP treatment.Conclusions SP has a growth regulatory property on cultivated bile duct fibroblast-like cells in vitro,suggesting that SP may involve in wound healing after bile duct injury by promoting wound fibroblast proliferation and inhibiting apoptosis and participate in pathological scar formation.

彩超联合CA199诊断肝内胆管结石合并肝内胆管细胞癌的价值

目的分析在诊断肝内胆管结石合并肝内胆管细胞癌时采取彩超联合CA199诊断方式的应用价值。方法抽取2017年7月—2022年12月我院肝胆胰外科收治的患者180例做为研究对象,并对其临床资料进行回顾性分析。以病理诊断结果为金标准进行分组,对照组共计150例,均为单纯肝内胆管结石患者,观察组共计30例,为肝内胆管结石合并肝内胆管细胞癌患者。所有患者均接受彩超检查、CA199检测,分析彩超联合CA199诊断肝内胆管结石合并肝内胆管细胞癌的诊断价值。结果对两组患者一般资料进行分析可见,观察组患者中男性占比高于对照组、有上腹部疼痛及高热症状的患者占比均低于对照组(均P<0.05);两组患者梗阻性黄疸、体重减轻、腹水、肝区叩痛以及肝外淋巴结肿大患者占比对比均无差异(均P>0.05)。彩超联合CA199诊断的准确度及特异度均高于单独应用彩超或单独应用CA199诊断(均P<0.05);彩超联合CA199诊断的敏感度与单独应用彩超或CA199诊断的敏感度对比均无差异(均P>0.05)。结论肝内胆管结石合并肝内胆管细胞癌诊断难度较高,易被误诊或漏诊,采取彩超联合CA199诊断的方式可显著提升诊断准确性,可为临床进行诊治提供较为可靠的依据。

智能水平旋转式细胞培养装置设计

目的:研制一种模块化、操作简单、易消毒、成本低、稳定性高的智能水平旋转式细胞培养装置,以用于管状支架内表面的细胞三维动态培养。方法:该装置由旋转培养模块、驱动模块、控制模块和控制软件组成,每个模块的外壳均由3D打印制作。其中,旋转培养模块由管状静电纺丝支架、细胞培养仓、磁耦合转子、聚丙烯管道等组成;驱动模块由N20减速电机、磁耦合转子组成;控制模块由ESP-8266芯片及印刷电路板组成;控制软件通过Blinker物联网平台开发,采用C++语言编程。采用该装置培养人肝内胆管上皮细胞,验证该装置的应用效果。结果:光镜和扫描电子显微镜图像显示,在管状静电纺丝支架表面形成了均匀连续的细胞层。结论:该智能水平旋转式细胞培养装置可实现管状静电纺丝支架内表面细胞的均匀生长,为管状支架上的细胞培养提供了有效平台。

lncRNA AL365181.2在胆管癌中的表达及生物学特性

目的探究长链非编码RNA(lncRNA)AL365181.2在胆管癌(cholangiocarcinoma,CHOL)中的表达水平及其对CHOL细胞功能学的影响。方法对肿瘤基因组图谱(TCGA)数据库中有关CHOL患者癌组织和癌旁组织的lncRNA AL365181.2表达情况的资料进行比较,对lncRNA AL365181.2表达水平与CHOL组织免疫细胞浸润情况的关系进行分析,并对lncRNA AL365181.2进行GSEA和KEGG富集分析。通过EdU实验、划痕实验、Transwell实验和裸鼠成瘤实验分别检测lncRNA AL365181.2对CHOL细胞增殖、迁移、侵袭和成瘤能力的影响。结果对TCGA数据库所涉资料的分析结果显示,与癌旁组织相比,CHOL组织中lncRNA AL365181.2显著高表达(Z=4.00,P<0.05),且lncRNA AL365181.2与CHOL组织的记忆T细胞及树突状细胞浸润水平显著相关(r=0.28、-0.34,P<0.05);GSEA分析结果显示lncRNA AL365181.2在CHOL细胞免疫调节相关通路中显著富集,KEGG富集分析结果显示lncRNA AL365181.2在细胞因子受体互作通路和血管壁细胞表面互作通路中显著富集。体内外实验结果显示,敲低lncRNA AL365181.2后,CHOL细胞的增殖、迁移、侵袭及体内成瘤能力均显著降低(t=4.23~173.34,P<0.05)。结论lncRNA AL365181.2在CHOL组织中较癌旁组织中高表达,并且表达水平与免疫细胞浸润程度密切相关;lncRNA AL365181.2的表达水平与CHOL细胞的增殖能力、迁移能力和侵袭能力密切相关。

蒿芩清胆汤调控枯否细胞M1/M2极化对原发性胆汁性胆管炎的机制研究

目的:基于枯否细胞M1/M2极化探讨蒿芩清胆汤对原发性胆汁性胆管炎(PBC)肝内胆管的保护作用及机制。方法:60只小鼠随机分为对照组、模型组、蒿芩清胆汤(HQQD)不同剂量组、熊去氧胆酸(UDCA)组,利用2-辛炔酸-牛血清白蛋白(2-OA-BSA)联合聚肌胞苷酸诱导的PBC小鼠模型,相应药物灌胃干预12周。苏木精伊红(HE)染色法观察肝组织病理学改变,全自动生化分析仪检测血清γ-谷氨酰转肽酶(γ-GT)、总胆汁酸(TBA)、总胆红素(TBil)、天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)改变,实时荧光定量(RT-PCR)检测一氧化氮合成酶(iNOS)、肿瘤坏死因子α(TNF-α)、甘露糖受体1(CD206)、白介素10(IL-10)mRNA的表达变化,蛋白免疫印记法(WB)检测iNOS、TNF-α、CD206、IL-10蛋白表达变化情况。结果:病理学结果显示,对照组小鼠肝组织门管区结构清晰,小叶间胆管周围无异常改变,模型组小鼠门管区可见多发炎症细胞聚集,炎症细胞聚集范围内可见肉芽肿、淋巴细胞浸润,HQQD高、中、低剂量组与UDCA组门管区炎症细胞数量较模型组减轻。生化分析结果显示,与对照组相比,模型组小鼠血清γ-GT、TBA、TBil、AST、ALT水平均不同程度升高;与模型组比较,UDCA组和HQQD高、中、低剂量组小鼠血清γ-GT、TBA、TBil、AST、ALT表达均明显下调。RT-PCR和WB结果显示,与对照组相比,模型组小鼠肝组织iNOS、TNF-αmRNA及蛋白表达水平明显上调,CD206、IL-10 mRNA及蛋白表达明显下调;与模型组比较,UDCA组和HQQD高、中、低剂量组iNOS、TNF-αmRNA及蛋白表达水平明显下调,CD206、IL-10 mRNA及蛋白表达水平明显上调。结论:蒿芩清胆汤可以使PBC模型小鼠枯否细胞M1极化减弱,M2极化增强,调控M1/M2极化状态,改善胆汁淤积及肝内胆管损伤。

术前肿瘤标志物检测对肝内胆管细胞癌淋巴结转移的预测价值分析

目的:分析术前肿瘤标志物检测对肝内胆管细胞癌(ICC)淋巴结转移的预测价值。方法:选取2021年1月—2023年12月海军军医大学第三附属医院收治的ICC患者78例作为研究对象。所有患者进行糖类抗原19-9(CA19-9)、糖类抗原242(CA242)、糖类抗原125(CA125)、癌胚抗原(CEA)、鳞状上皮癌细胞抗原(SCC)检测,并以手术探查结果及术后病理检查结果为“金标准”判定是否存在淋巴结转移。比较不同转移情况患者肿瘤标志物指标水平及阳性率,采用受试者操作特征(ROC)曲线分析术前肿瘤标志物检测对于淋巴结转移的预测价值。结果:经“金标准”评估,78例肝内胆管癌患者分为转移组(n=31)与未转移组(n=47)。转移组CA19-9、CA242、CA125指标水平及阳性率均高于未转移组(P<0.05);两组CEA、SCC指标水平及阳性率比较,差异均无统计学意义(P>0.05)。ROC曲线分析结果显示,CA19-9、CA242、CA125用于预测ICC淋巴结转移的AUC分别为0.776、0.795、0.826。其中CA125预测ICC淋巴结转移的灵敏度最高,为87.10%。结论:术前CA19-9、CA242、CA125指标检测在ICC淋巴结转移的预测中具有较高的应用价值。

微RNA-196a-1-3p靶向Ras响应元件结合蛋白调控胆管癌细胞增殖的机制研究

目的探讨转化生长因子β(TGF-β)调控人胆管癌细胞系RBE细胞增殖的关键微RNA(miRNA)及其潜在的机制。方法该研究起止时间为2020年1月至2022年1月。磷酸盐缓冲液(PBS)处理为对照组,TGF-β处理为TGF-β组,TGF-β抗体处理为抗体组。检测三组RBE细胞的增殖水平。miRNA高通量测序检测三组RBE细胞的miRNA调控变化,并进行miRNA模拟物过表达筛选鉴定受TGF-β调控的影响RBE细胞增殖水平的关键miRNA。miRNA数据库(miRDB)在线分析miRNA的潜在底物,并通过小干扰RNA(siRNA)敲低筛选鉴定影响RBE细胞增殖水平的关键底物。结果相比于对照组,TGF-β组RBE细胞的增殖水平上升(1.62±0.07比2.35±0.09,P<0.05),抗体组RBE细胞的增殖水平下降(1.62±0.07比1.11±0.08,P<0.05)。过表达微RNA-196a-1-3p(miR-196a-1-3p)时,RBE细胞的增殖水平下降(P<0.05)。敲低Ras响应元件结合蛋白(RREB1)时,RBE细胞的增殖水平下降(P<0.05)。过表达miR-196a-1-3p后,RBE细胞中RREB1的信使RNA(mRNA)和蛋白水平下降(P<0.05)。敲低miR-196a-1-3p后,RBE细胞中RREB1与SMAD家族蛋白3(SMAD3)的相互作用增加。敲低SMAD3后,RBE细胞的增殖水平下降(P<0.05)。与仅敲低SMAD3相比,敲低SMAD3的同时过表达RREB1的RBE细胞的增殖水平无显著变化,并且同时敲低SMAD3和miR-196a-1-3p的RBE细胞的增殖水平无显著变化。结论TGF-β能够通过miR-196a-1-3p/RREB1/SMAD3轴促进RBE细胞增殖;miR-196a-1-3p和RREB1可作为潜在的治疗胆管癌的靶标,为针对该靶标的新药研发奠定了基础。

热化疗对人胆管癌细胞增殖和凋亡的影响

目的探讨表阿霉素(EADM)热化疗对人胆管癌细胞(QBC939)的增殖和凋亡的作用。方法以体外培养的人胆管癌细胞株为研究对象,采用水浴加热法进行体外细胞毒实验(MTT)、透射电镜观察及流式细胞仪检测,观察热疗、化疗、热化疗对人胆管癌细胞的生长抑制和凋亡的影响。结果42℃以上单纯热疗对QBC939细胞有明显杀伤作用(P<0.01),42℃以上热化疗对QBC939细胞有明显的协同或相加作用(P<0.01)。透射电镜和流式细胞术均观察到热疗、化疗、热化疗诱导细胞凋亡的作用(P<0.01);热化疗有协同作用(P<0.01)。结论热疗、化疗、热化疗均抑制QBC939细胞增殖;热疗、化疗、热化疗诱导细胞凋亡为其抗癌机制之一;热疗提高化疗药物的细胞毒作用。