p27kip1基因在人胆管癌细胞化疗增敏效应中的作用

目的 探讨外源性p27^kip1在人胆管癌细胞系QBC939中高表达对胆管癌细胞化疗敏感性的影响。方法 通过携带人p27^kip1基因的重组腺病毒载体Ad-p27mt转染人胆管癌细胞系QBC939。经逆转录-聚合酶链反应（RT-PCR）、Western blot检测p27^kip1在胆管癌细胞中的表达；经MTT比色法及流式细胞术检测5-氟尿嘧啶（5-FU）、丝裂霉素（MMC）、环磷酰胺（CTX）对转染p27^kip1前后的QBC939细胞生长抑制及凋亡的影响。结果 Ad-p27mt转染后5-FU、MMC和CTX对QBC939细胞生长抑制率，分别由转染前的41．89％、45．59％和38．91％显著增加为56．15％、55．65％和51．69％；凋亡率也由13．76％、11．76％和10．46％，明显升高为41．39％、35．94％、34．46％，差异均有显著性意义（均P〈0．05）。结论 p27^kip1在QBC939细胞中高表达，能显著增强胆管癌细胞对化疗药物的敏感性，为基因联合化疗药物治疗胆管癌提供了参考依据。     

Rb反义寡核苷酸对人结肠癌细胞凋亡及化疗药敏的影响

目的　观察Rb反义寡核苷酸 (AS ODN)对人结肠癌(HT 2 9)细胞凋亡及化疗药物敏感性的影响。方法　人工合成RbAS ODN经脂质体 (Lip)包裹后作用HT 2 9细胞。采用免疫组化、WesternBlot检测转染前后Rb蛋白表达状况 ,流式细胞术测定HT 2 9细胞转染前后 5 Fu诱导凋亡程度 ,MTT法观察化疗药敏的变化。结果　经RbAS ODN处理的HT 2 9细胞Rb蛋白表达量明显下降 ,RbAS ODN处理的HT 2 9细胞加 5 Fu作用后与对照组比较 ,细胞凋亡率明显下降 ,化疗药物敏感性降低。结论　RbAS ODN阳离子脂质体转染具有抑制化疗药诱导大肠癌HT 2 9细胞凋亡的作用及降低化疗药物敏感性作用。     

宫颈鳞状细胞癌ATP-TCA法体外药敏试验与耐药相关蛋白表达的关系

目的:探讨代表不同耐药机制的P糖蛋白(P-gp)、谷胱甘肽S-转移酶(GST-π)、DNA拓扑异构酶Ⅱ(TopoⅡ)、胸苷酸合成酶(TS)在原发宫颈鳞癌组织中的表达及其与临床病理参数的关系;分析三磷酸腺苷生物荧光法体外药敏试验(ATP-TCA)结果与耐药蛋白表达的关系。方法:收集32例宫颈鳞癌患者的新鲜癌组织制成单细胞悬液,采用ATP-TCA法成功检测上述细胞对11种化疗药物(共16种组合)的体外敏感性;采用EnVision二步法免疫组化检测32例宫颈鳞癌组织中P-gp、GST-π、TopoⅡ、TS蛋白的表达,并分析其与临床分期、肿瘤分化程度的关系。结果:耐药蛋白P-gp、GST-π、TopoⅡ、TS在宫颈鳞癌组织中的表达与临床分期、肿瘤分化程度无相关性;耐药蛋白P-gp表达与体外PTX+CBP耐药正相关(P=0.040);耐药蛋白TS表达与体外5-FU、5-FU+MMC、5-FU+DDP耐药正相关(P=0.015,0.012,0.006);TopoⅡ,GST-π蛋白表达与宫颈鳞癌体外耐药均无相关性。结论:体外药敏试验联合免疫组化检测,有可能用于宫颈鳞癌化疗前药敏预测。     

DCLK1 对结直肠癌细胞化疗敏感性的影响及机制

目的:探讨双肾上腺皮质激素样激酶1（DCLK1）对结直肠癌耐药细胞化疗敏感性的影响及机制。方法:采用实时定量PCR和Western blot方法检测人结直肠癌细胞系HT29、SW480及其奥沙利铂（OXA）耐药株HT29/OXA和SW480/OXA中DCLK1的表达。采用siRNA干扰HT29/OXA和SW480/OXA细胞中DCLK1的表达,并用不同浓度OXA干预细胞,CCK-8法检测细胞增殖及其对OXA的敏感性,Annexin V-FITC/PI双染法检测细胞凋亡,Caspase-3活性检测试剂盒检测细胞的Caspase-3活性,Western blot检测细胞中DCLK1、Bax、Bcl-2、多药耐药蛋白1（MDR1）、P-糖蛋白（P-gp）等蛋白的表达。结果:与亲本细胞系比较,DCLK1在HT29/OXA和SW480/OXA细胞中的mRNA和蛋白表达水平均显著上调（P<0.05）。转染DCLK1 siRNA（si-DCLK1）后,HT29/OXA和SW480/OXA细胞的增殖活性均显著降低（P<0.05）,且它们对OXA的敏感性显著增加（P<0.05）;转染si-DCLK1后,HT29/OXA和SW480/OXA细胞的凋亡显著增加（P<0.05）,伴随Caspase-3活性上调（P<0.05）,Bax蛋白表达上调（P<0.05）,Bcl-2蛋白表达下调（P<0.05）;转染si-DCLK1后,HT29/OXA和SW480/OXA细胞中MDR1和P-gp的蛋白表达均显著下调（P<0.05）。结论:DCLK1可能通过调控Bax/Bcl-2以及MDR1和P-gp的表达影响结直肠癌细胞的化疗敏感性。     

三磷酸腺苷-肿瘤体外药敏试验对宫颈癌临床化学治疗的指导作用

目的：研究三磷酸腺苷-肿瘤体外药敏试验（ATP-tumor chemosensitivity assay,ATP-TCA）对宫颈癌患者化学治疗（化疗）方案选择的指导性作用。方法：选择2007年7月至2009年10月宫颈癌住院患者72例,将其随机分为药敏组（35例）和对照组（37）例。采用ATP-TCA法检测药敏组宫颈癌组织对6种联合化疗方案的敏感性。药敏组患者根据药敏结果选择化疗方案,对照组根据临床经验制定化疗方案。随访3年,比较药敏组与对照组1年复发率和3年生存率。结果：3s例药敏组中有32例成功进行了ATP-TCA检测;体外有效例数分别为紫杉醇＋卡铂20例、紫杉醇＋奥沙利铂18例、博莱霉素＋异环磷酰胺＋顺铂17例、博莱霉素＋长春新碱＋顺铂18例、氟尿嘧啶＋顺铂17例、吉西他滨＋顺铂21例。1年复发例数药敏组与对照组比较差异无统计学意义（P〉0．05）;3年生存情况药敏组好于对照组（P〈0．05）。结论：ATP-TCA法可以为宫颈癌患者筛选出敏感、有效、个体化的化疗方案,值得临床推广。     

BNIP3在胰腺癌中的表达及其与术后吉西他滨化疗敏感性的关系

目的 探究胰腺癌Bcl-2/腺病毒E1B-19k相关蛋白3(Bcl-2/E1B-19k-interacting protein 3,BNIP3)的蛋白表达水平及其与术后吉西他滨化疗敏感性的关系。方法 回顾性分析2016年7月至2021年8月内蒙古医科大学附属医院58例胰腺癌根治术,并在术后规律应用吉西他滨辅助化疗或以吉西他滨为主的联合化疗患者的临床资料,选取胰腺癌术后组织切片,对其进行BNIP3免疫组织化学检测,通过实验染色强度及范围进行评分,并分析其与胰腺癌患者预后的关系。结果 与BNIP3低表达组比较,BNIP3高表达组的总生存时间(OS)[13(95%CI 11.378～14.622)个月 vs 18(95%CI 16.592～19.408)个月]和无病生存时间(DFS)[7(95%CI 4.082～9.938)个月 vs 14(95%CI 12.936～15.064)个月]均降低(P<0.05)。BNIP3的蛋白表达水平及淋巴转移情况是OS的独立预后因素,而BNIP3的蛋白表达水平及分化程度是DFS的独立预后因素。结论 胰腺癌组织中BNIP3的蛋白表达水平可能与应用吉西他...     

Short‐term intermittent hypoxia reduces the severity of acute mountain sickness

Intermittent hypoxia (IH) is a promising approach to induce acclimatization and hence lower the risk of developing acute mountain sickness (AMS). We hypothesized that a short‐term IH protocol in normobaric hypoxia (7 × 1 h to 4500 m) effectively increases the hypoxic ventilatory response (HVR) and reduces the incidence and severity of AMS. Therefore, 26 men (25.5 ± 4.4 years), assigned in a double‐blinded fashion to the hypoxia group (HG) or placebo group (PG), spent 8 h at 5300 m before (PRE) and 2 days after cessation of the IH protocol (POST). Measurements included the evaluation of the Lake Louise Score (LLS) and the HVR. The severity of AMS decreased from PRE to POST in the HG (from 6.0 ± 2.7 at PRE to 4.1 ± 2.1 at POST), whereas the LLS in the PG stayed high (from 5.7 ± 2.9 to 5.5 ± 2.8, respectively). The HVR in the HG increased from 0.73 ± 0.4 L/min/% at PRE to 1.10 ± 0.5 L/min/% at POST and did not increase in the PG. The reduction of the LLS was inversely related to the changes in the HVR (r = ?0.434), but the AMS incidence was not different between the HG and the PG at POST. In conclusion, short‐term IH reduced the severity of AMS development during a subsequent 8‐h exposure to normobaric hypoxia.     

LRIG1 Enhances Chemosensitivity by Modulating BCL-2 Expression and Receptor Tyrosine Kinase Signaling in Glioma Cells

Purpose

Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) are an inhibitor of receptor tyrosine kinases (RTKs) that was discovered in recent years, and many studies showed that LRIG1 is a tumor suppressor gene and may be related to tumor drug resistance. In this study, we explored whether LRIG1 protein expression can improve the chemosensitivity of glioma cells and what was its mechanism.

Materials and Methods

We collected 93 cases of glioma tissues and detected the expression of LRIG1 and BCL-2. We constructed a multidrug resistance cell line U251/multidrug resistance (MDR) and examined the change of LRIG1 and BCL-2 at mRNA and protein expression levels. LRIG1 expression was upregulated in U251/MDR cells and we detected the change of multidrug resistance. Meanwhile, we changed the expression of LRIG1 and BCL-2 and explored the relationship between LRIG1 and BCL-2. Finally, we also explored the relationship between LRIG1 and RTKs.

Results

LRIG1 was negatively correlated with BCL-2 expression in glioma tissue and U251/MDR cells, and upregulation of LRIG1 can enhance chemosensitivity and inhibit BCL-2 expression. Furthermore, LRIG1 was negatively correlated with RTKs in U251/MDR cells.

Conclusion

These results demonstrated that LRIG1 can improve chemosensitivity by modulating BCL-2 expression and RTK signaling in glioma cells.     

Multi-disciplinary treatment of a rare pelvic cavity ependymoma

Ependymomas usually develop from neuroectodermal organs. Here, we present an ependymoma arising from the pelvic cavity. A 27-year-old Korean female was admitted to the hospital with a sensation of abdominal fullness. Imaging studies revealed a huge heterogeneous nodular mass in the pelvis and lower abdomen. Laparotomy showed that two large masses with multiple nodules were located between the uterus and rectum and uterus and bladder, respectively. Histologically, the tumor was characterized by compact columnar neoplastic cells divided by fibrovascular septae. The neoplastic cells formed true ependymal rosettes and perivascular pseudorosettes. Immunohistochemical staining showed a strong positive reaction for glial fibrillary acidic protein (GFAP) and vimentin and a partial positive reaction for S100 and EMA. The tumor was thus diagnosed as an ependymoma arising from the pelvic cavity. The patient was treated with a debulking operation and chemotherapy based upon the in vitro chemosensitivity test results. The patient was free of cancer for 4 years following surgery. This is a rare case of extraneural ependymoma for which an in vitro chemosensitivity test was critical in determining the multidisciplinary approach for treatment.     

RNA干扰抑制EZH2基因表达对人胶质瘤细胞凋亡及对卡莫司汀敏感性的影响

目的观察RNA干扰技术抑制EZH2基因表达对人胶质瘤U251细胞凋亡及其对卡莫司汀敏感性的影响。方法针对EZH2 mRNA序列设计合成小干扰RNA（siRNA）的DNA模板,构建pRNAT-EZH2重组表达载体,并用其转染人胶质瘤U251细胞。用RT-PCR法检测pRNAT-EZH2转染前后U251细胞中内源性EZH2表达;激光共聚焦显微镜观察转染后及加入卡莫司汀后U251细胞凋亡情况;MTT法检测转染前后U251细胞对化疗药物卡莫司汀敏感性;用分光光度法检测转染前后以及加入卡莫司汀后U251细胞中Caspase-3活性。结果成功构建pRNAT-EZH2重组质粒,并成功转染U251细胞。pRNAT-EZH2重组质粒转染后U251细胞中EZH2 mRNA表达量降低（P〈0.01）;卡莫司汀组及卡莫司汀加pRNAT-EZH2组U251细胞凋亡明显;与卡莫司汀联合时,pRNAT-EZH2组U251细胞生长抑制率明显增高（P〈0.01）;Caspase-3活性明显高于空白对照组和阴性对照组（P均〈0.01）。结论pRNAT-EZH2可抑制EZH2在人胶质瘤U251细胞中的表达,并增强U251细胞对卡莫司汀敏感性。