人胆囊癌细胞系GBC-SD中侧群细胞的耐药性研究

目的 研究人胆囊细胞系GBC-SD中侧群细胞(side population cells,SP)的耐药特性,并探讨其耐药机制.方法 利用流式细胞术分选SP、非SP细胞,采用MTT检测2种细胞亚群对5种化疗药物吉西他滨、顺铂、5-氟尿嘧啶、表阿霉素、米托蒽醌的药物敏感性;利用流式细胞术检测吉西他滨处理的人胆囊癌细胞系GBC-SD中SP细胞比例的变化,并通过RT-PCR、Western印迹检测SP、非SP细胞亚群中ABCG2 mRNA和蛋白的表达.结果 吉西他滨、顺铂、5-氟尿嘧啶、米托蒽醌以对胆囊癌细胞系GBC-SD的IC50浓度分别作用SP、非SP细胞亚群1 d后,SP细胞的增殖能力高于非SP细胞,两组之间的差异具有统计学意义(P<0.05),而表阿霉素处理水平的两组间差异无统计学意义(P>0.05);人胆囊癌细胞系GBC-SD经过吉西他滨处理3周后,SP细胞的比例显著升高(8.02%±0.13%比0.62%±0.08%,P<0.05),且SP细胞明显高表达ABCG2基因.结论 人胆囊癌细胞系GBC-SD中SP细胞具有类似干细胞的耐药特性,耐药基因ABCG2高表达是其耐药的重要机制.

Abstract：

Objective To investigate the drug resistance of side population cells in human gallbladder cancer cell line GBC-SD and explore its mechanism. Methods Drug sensitivity assays of 5chemotherapeutic agents were performed on side population cells (SP) and non-SP cells of GBC-SD.GBC-SD was cultured and then treated with the chemotherapeutic agent gemcitabine. The frequency of SP by FACS was measured. RT-PCR and Western blotting were used to detect the expression of AB-CG2 in both the SP and the corresponding non-SP subsets. Results After 1 d treatment with 4 chemotherapeutic agents (gemcitabine, cisplatin, 5-fluorouracil and mitoxantrone) in IC50 concentration to GBC-SD cell line, the reproductive ability of SP was higher than that of non-SP (P<0.05). However, statistical significance was not achieved when compared with epirubicin (P>0.05). The percentage of SP in GBC-SD treated with chemotherapeutic agent gemcitabine after 3 weeks was sharply elevated by FACS (8.02% ±0.13% vs 0.62% ±0.08%, P<0.05), and the expression of ABCG2mRNA and protein were increased in SP as compared with non-SP. Conclusion SP from human gallbladder cancer cell line GBC-SD, like stem cell, showed a heighten resistance to drugs. Increased expression of ABCG2 was largely responsible for the multi-drug resistance.     

人胆囊癌细胞系GBC-SD侧群细胞致瘤特性的实验研究

目的 研究人胆囊癌细胞系GBC-SD侧群细胞(side population cells,SP)的致瘤特性。方法 利用流式细胞术从GBC-SD中分选SP、非SP细胞(non-SP),分别进行软琼脂克隆形成实验和非肥胖性糖尿病联合免疫缺陷(NOD/SCID)小鼠的移植瘤形成实验。通过流式细胞术检测5例人胆囊癌组织中SP细胞的比例。结果人胆囊癌细胞系GBC-SD中存在SP细胞,其比例约为0.87％;SP细胞的克隆形成率高于non-SP细胞（14.74％±3.53％比5.17％±1.05％,t=2.75,P＜0.05）。NOD/SCID小鼠移植瘤实验表明,5×103个SP细胞可成瘤(4/7),而non-SP细胞成瘤则需要1×105个细胞(1/7),且来源于SP细胞亚群的NOD/SCID小鼠移植瘤中存在SP、non-SP细胞。SP细胞存在于人胆囊癌组织中,其范围为0.27％～2.3％。结论来源于GBC-SD的SP细胞亚群具有类似肿瘤十细胞的高致瘤性。     

头颈部CTA对颅内破裂动脉瘤早期诊断价值研究

目的 以数字减影血管造影(digital subtraction angiography, DSA)为标准,探讨头颈部CT血管成像(computed tomography angiography, CTA)对颅内破裂动脉瘤病变部位、大小、形态特征早期诊断价值。方法 回顾性分析2018年1月—2021年10月本院收治的发病在5天内因颅内动脉瘤破裂致蛛网膜下腔出血(subarachnoid hemorrhage, SAH)行CTA检查并行DSA检查的患者61例的临床资料。比较分析这两种方法测量破裂的动脉瘤部位、形态参数,分析灵敏度、特异度、阳性预测值、阴性预测值、颅内动脉瘤的直径、瘤颈宽度,动脉瘤颈部和母体动脉的三维关系、图像质量优良率比较。结果 CTA和DSA检查破裂动脉瘤部位、动脉瘤颈部和母体动脉的三维关系比较、瘤体直径及瘤颈宽度比较差异无统计学意义(P>0.05)。CTA与DSA检查的图像质量优良率之间的差异无统计学意义(P>0.05)。对于直径<4 mm的破裂颅内动脉瘤,CTA的灵敏度为90.0%,特异度为71.43%,阳性预测值为90.0%,阴性预测值为71.4...