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组织培养药物敏感性检测技术

组织培养药物敏感性检测技术. Histoculture Drug Response Assay (HDRA). HDRA 技术介绍. HDRA 技术由 Hoffman 等人于80年代末期建立, 是一种基于非分散性组织的药物敏感性检测技术,该技术将微小组织培养于一种天然胶原海绵基质上,同时加入抗癌药物对其作用一定的时间,细胞活性终点评价采用 MTT 还原法,结果较为直观可靠. 应用癌种.

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组织培养药物敏感性检测技术

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  1. 组织培养药物敏感性检测技术 Histoculture Drug Response Assay (HDRA)

  2. HDRA技术介绍 • HDRA技术由Hoffman等人于80年代末期建立, 是一种基于非分散性组织的药物敏感性检测技术,该技术将微小组织培养于一种天然胶原海绵基质上,同时加入抗癌药物对其作用一定的时间,细胞活性终点评价采用MTT还原法,结果较为直观可靠.

  3. 应用癌种 • 胃癌 肠癌 卵巢癌 乳腺癌 肉瘤 肺癌 泌尿系肿瘤 头颈部肿瘤

  4. HDRA技术流程 • 1.肿瘤组织预处理:去脂肪,纤维以及血污. • 2.肿瘤组织处理:清洗,眼科镊子和小剪刀进行剪切,使组 • 织块大小约为1-2mm. • 3.将3块组织块放置于约1cm2的胶原海绵小块上,然后放 • 置于预加有培养基的24孔板中,培养过夜. • 4.加入含药物的培养基,每个浓度2个平行孔. • 5.继续培养3天后,加入MTT,孵育4h,然后提取还原的MTT,测 • 定570nm的OD值. • 6.药物处理组和对照组进行比较,计算抑制率,评价药物杀伤.

  5. 肿瘤组织的处理

  6. 培养中的肿瘤组织(一) 培养孔 培养基液面 肿瘤组织 胶原海绵

  7. 培养中的肿瘤组织(二)

  8. HDRA培养肿瘤组织形态结构

  9. MTT还原法测定细胞活力

  10. 结果评价 • 加药处理组(T) • 空白对照组(C) • 抑制率(I)=T/C • 敏感药物:I>50%

  11. HDRA技术特点 • 1.肿瘤组织不需要进行机械/酶学分散,保持其结构及形 • 态,减少操作造成的细胞损失. • 2.模拟体内药物对癌组织的作用,评价客观,同时避免了 • 仅进行癌细胞评价的片面性. • 3.该技术采用天然胶原海绵作为培养基质,同时培养的 • 组织接近液面,促进气液交换,保证癌细胞的生长增殖. • 4.具有较高的标本可评价率,据报道为90~100%.

  12. HDRA技术历史和现状 • 1.Hoffman等人于80年代末期建立基于胶原海绵的立体 • 组织培养技术,后应用于原代瘤组织体外药敏检测,至 • 今已有10余年的历史. • 2.目前该技术主要在日本和美国进行大量的临床应用 • 研究, 公开发表文献50余篇. • 3.2002年Singh等人在头颈部肿瘤中的研究表明,该技术 • 能够评价患者预后,从而指导肿瘤化学治疗.

  13. HDRA临床相关性研究

  14. HDRA临床应用研究

  15. 乳腺癌

  16. 小细胞肺癌

  17. 肠癌

  18. 肺癌

  19. 胰腺癌

  20. HDRA与胃肠癌生存预后 The survival rate of 10 patients whose tumors were sensitive to either mitomycin C and/or 5-fluorouracil in the assay was significantly (P < 0.005) better than that of 22 patients whose tumors were shown to be insensitive to both drugs. Twenty-nine patients with stage III and IV colorectal cancer without remaining measurable tumor lesions after surgery were treated with fluoropyrimidines adjuvantly. The recurrence-free survival rate of 7 patients whose tumors were sensitive to 5-fluorouracil in the assay was significantly (P < 0.05) better than that of 22 patients whose tumors were insensitive.

  21. Kaplan miere 生存曲线

  22. HDRA检测与头颈癌生存预后 • Resistance to 5-fluorouracil was present in 13 cases (32%), to cisplatinum in 13 cases (32%), and to both agents in 11 cases (27%). The 2-year cause-specific survival was significantly greater for patients sensitive to 5-fluorouracil (85% vs 64%; p =.04), cisplatinum (86% vs 64%; p =.05), or both agents (85% vs 63%; p =.01).

  23. Kaplan miere 生存曲线

  24. 结论 • HDRA技术是一种基于非分散性组织的药物敏感性检测技术,该技术保持肿瘤组织基本形态结构,模拟药物对肿瘤组织的作用,同时避免了由于组织分散造成的细胞损伤和丢失.国外研究结果表明,该技术准确,可靠,与临床疗效具有较好的相关性,目前已在美国和日本进入临床应用.近年来的研究表明,该技术能够反映肿瘤患者生存预后,对于指导个体化治疗意义重大.

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