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中华普通外科学文献(电子版) ›› 2021, Vol. 15 ›› Issue (06) : 418 -424. doi: 10.3877/cma.j.issn.1674-0793.2021.06.004

论著

不同处理因素对结直肠癌类器官奥沙利铂敏感性测试结果的影响研究
李武国1, 陈伟2, 苏乔1,(), 李雯雯1, 赵广银1, 杨宇童1, 刘长琳1   
  1. 1. 510080 广州,中山大学附属第一医院动物实验中心
    2. 518107 深圳,中山大学附属第七医院病理科
  • 收稿日期:2021-07-07 出版日期:2021-12-01
  • 通信作者: 苏乔
  • 基金资助:
    广东省科技计划项目(2018B030317001、2019A030317005); 广东省结直肠盆底疾病研究重点实验室开放基金项目(2018-01)

Effect of different processing factors on oxaliplatin sensitivity test results of colorectal cancer organoids

Wuguo Li1, Wei Chen2, Qiao Su1,(), Wenwen Li1, Guangyin Zhao1, Yutong Yang1, Changlin Liu1   

  1. 1. Animal Experiment Center, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China
    2. Department of Pathology, the Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen 518107, China
  • Received:2021-07-07 Published:2021-12-01
  • Corresponding author: Qiao Su
引用本文:

李武国, 陈伟, 苏乔, 李雯雯, 赵广银, 杨宇童, 刘长琳. 不同处理因素对结直肠癌类器官奥沙利铂敏感性测试结果的影响研究[J/OL]. 中华普通外科学文献(电子版), 2021, 15(06): 418-424.

Wuguo Li, Wei Chen, Qiao Su, Wenwen Li, Guangyin Zhao, Yutong Yang, Changlin Liu. Effect of different processing factors on oxaliplatin sensitivity test results of colorectal cancer organoids[J/OL]. Chinese Archives of General Surgery(Electronic Edition), 2021, 15(06): 418-424.

目的

选用结直肠癌临床常用药物奥沙利铂,探究不同处理因素对结直肠癌类器官药物敏感性测试结果的影响,为肿瘤患者个体化精准治疗方案的筛选提供测试条件参考。

方法

采用不同细胞密度、给药前不同细胞培养时间、不同药物作用时间对2例结直肠癌类器官CRC51、CRC67进行处理,设计100、10、1、0.1、0.01 μmol/L 5个剂量的奥沙利铂对2例结直肠癌类器官进行作用,CCK-8检测细胞增殖活力,根据细胞存活率(T/C)比较分析不同处理因素对结直肠癌类器官药物敏感性测试结果的影响。

结果

①在细胞密度的选择上,每孔接种8 000个和4 000个细胞时,100 μmol/L的奥沙利铂均对类器官CRC51、CRC67的生长产生显著的抑制作用,两者的抑制作用差异无统计学意义。而每孔接种2 000个细胞时,100 μmol/L的奥沙利铂对类器官CRC51、CRC67的生长抑制作用均显著变弱(与每孔接种8 000个和4 000个细胞相比,P<0.05),药效评价结果变为体外无效。②在给药前细胞培养时间的选择上,没有获得规律性的研究结果,但均T/C>50%。③在药物作用时间的选择上,100 μmol/L的奥沙利铂对类器官CRC51、CRC67作用7 d比作用3 d获得更加显著的生长抑制作用(P<0.05)。

结论

不同细胞密度、给药前不同细胞培养时间、不同药物作用时间对结直肠癌类器官药物敏感性测试结果均会产生影响。选择的实验条件不同,可能会得出相反的药效评价结果。

Objective

To explore the effects of different processing factors on oxaliplatin sensitivity test results of colorectal cancer organoids and provide reference for the individualized precision treatment for tumor patients.

Methods

Two cases of colorectal cancer organoids were processed with different cell density, different cell culture time prior to administration and different drug action time. Five doses of oxaliplatin were designed to act on the two cases of colorectal cancer organoids. CCK-8 was used to detect cell proliferation activity. The effects of different processing factors on drug sensitivity test results of colorectal cancer organoids were analyzed according to cell survival rate.

Results

In the selection of cell density, 100 μmol/L oxaliplatin could significantly inhibit the growth of colorectal cancer organoids in both cases when inoculated with 8 000 and 4 000 cells per well, and there was no significant difference between the two groups. When inoculated with 2 000 cells per well, the growth inhibitions of 100 μmol/L oxaliplatin on colorectal cancer organoids in both cases were significantly weakened (compared with 8 000 and 4 000 cells per well, P<0.05), and the results of efficacy evaluation became invalid in vitro. No regular results were obtained on the time of cell culture prior to administration. In the choice of time of action, 100 μmol/L oxaliplatin showed more significant growth inhibition effect on the two colorectal cancer organoids after being treated for 7 days than 3 days (P<0.05).

Conclusions

Different cell density, different cell culture time prior to administration and different drug action time show influence on the drug sensitivity test results of colorectal cancer organoids. Different experimental conditions may lead to opposite efficacy evaluation results.

图1 不同细胞密度(8 000、4 000、2 000个/孔)对结直肠癌类器官药物敏感性测试结果的影响 A、B分别为CRC51各组T/C百分比柱状图、百分比-对数浓度曲线图;C、F分别为CRC51、CRC67各组结直肠癌类器官处理后形态代表图(×100);D、E分别为CRC67各组T/C百分比柱状图、百分比-对数浓度曲线图;***P<0.001,** P<0.01,* P<0.05
图2 在相同细胞密度、奥沙利铂作用3 d条件下,给药前不同细胞培养时间(4 h、1 d)对结直肠癌类器官药物敏感性测试结果的影响 A、B分别为CRC51各组T/C百分比柱状图、百分比-对数浓度曲线图;C、F分别为CRC51、CRC67各组结直肠癌类器官处理后形态代表图(×100);D、E分别为CRC67各组T/C百分比柱状图、百分比-对数浓度曲线图;* P<0.05
图3 在相同细胞密度、奥沙利铂作用7 d条件下,给药前不同细胞培养时间(4 h、1 d)对结直肠癌类器官药物敏感性测试结果的影响 A、B分别为CRC51各组T/C百分比柱状图、百分比-对数浓度曲线图;C、F分别为CRC51、CRC67各组结直肠癌类器官处理后形态代表图(×100);D、E分别为CRC67各组T/C百分比柱状图、百分比-对数浓度曲线图;***P<0.001,**P<0.01
图4 在相同细胞密度、给药前细胞培养4 h条件下,不同药物作用时间(3、7 d)对结直肠癌类器官药物敏感性测试结果的影响 A、B分别为CRC51各组T/C百分比柱状图、百分比-对数浓度曲线图;C、F分别为CRC51、CRC67各组结直肠癌类器官处理后形态代表图(×100);D、E分别为CRC67各组T/C百分比柱状图、百分比-对数浓度曲线图;***P<0.001
图5 在相同细胞密度、给药前细胞培养1 d条件下,不同药物作用时间(3、7 d)对结直肠癌类器官药物敏感性测试结果的影响 A、B分别为CRC51各组T/C百分比柱状图、百分比-对数浓度曲线图;C、F分别为CRC51、CRC67各组结直肠癌类器官处理后形态代表图(×100);D、E分别为CRC67各组T/C百分比柱状图、百分比-对数浓度曲线图;**P<0.01
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