An Application Study Of The CD-DST(Collagen Gel Droplet Embedded Cultured Drug Sensitivity Testing) For Head And Neck Cancer

Objective:Of the substantial anticancer drug sensitivity testing methods, CD-DST is an advanced method that overcame several defects in the existing methods. This thesis aims to establish in vitro tumor response testing for head and neck cancer and to explore the value of application. The primary cultured tumor cells and the NPC cell lines CNE-1and CNE-1/taxol were embedded in collagen gel droplet to detect the sensitivity of the respective chemotherapy drugs (DOC、PAC、DDP and5-FU) and so as to make a comparative study thereof.Methods:1. A total of46fresh surgical specimens obtained from head and neck cancer patients from October2013to February2014at the Third XiangYa Hospital were collected. After the specimens had been processed, the cells were transferred into the collagen gel-coated flasks.2. Collect the primary tumor cells and the NPC cell lines CNE-1and CNE-1/taxol, count the number, then embedded the cells in the collagen gel droplet inoculated in6-well plate.24h later, the anticancer drugs (DOC、PAC、DDP and5-FU) were added and incubated for another24h. After removal of the medium containing the drugs, each well was rinsed twice, overlaid with serum-free medium, and incubated for a further5-7days. At the end of the incubation, neutral red was added to each well and then fixed with formalin.3. In-vitro sensitivity was expressed as the percentage T/C ratio, where T was stand for the image optical density (OD) of the treated group and C that of the control group:a T/C ratio of50%or less was regarded as demonstrating in vitro sensitivity.Results:1. The primary tumor cells we cultured shows its deeper nuclear staining, different sizes in each, clumps scattered and some embedded tightly. In collagen gel droplet embedded culture, the cell grew with a three-dimensional spherical morphology, mimicking their in-vivo characteristics.2. After addition of anticancer drugs, part of the primary tumor cells has disintegrated just as vacuoles. Nevertheless, the NPC cell lines CNE-1and CNE-1/taxol did not obviously changed.3. The overall evaluable rate was67.39%, including NPC (20/23)86.96%, tongue cancer (4/7)57.14%, laryngeal carcinoma (3/9)33.33%, hypopharyngeal carinoma (3/6)50%.4. The single drug efficiency from high to low in the order is DOC (45.16%), PAC (41.96%), DDP (29.03%),5-FU (16.13%).Conclusion:1. CD-DST has many advantages including a higher success rate (67.39%), the less need for only the number of cells for the test (3×103cells/drop), and a better correlation between in-vitro and in-vivo results. Therefore, it deserves some reference value in formulating personalized chemotherapy regimens.2. Of the HNC specimens, the success rate of NPC is the highest, and that of laryngeal carcinoma and that of hypopharyngeal carcinoma remain low, which might be related with the contamination and biological characteristics.3. CD-DST may not apply to the anticancer drug sensitivity testing for the cell lines.