Description:
<jats:title>Abstract</jats:title>
<jats:p>For diversely heterogeneous diseases like cancer, a major challenge in the development of new drugs is using appropriate in vitro testing platform. Although high throughput screening of new compounds requires a large panel of genetically well characterized cell lines, their numbers are far less than requirement of biophrama. On top of this the translational potential of cell line panel is limited because most of the commercially available cell lines have been generated from metastatic site and highly transformed, therefore do not represent the true state of the disease. In this context, we have developed two early passage patient derived primary colorectal cancer (CRC) cell lines. Using this improved in vitro testing system we tested the efficacy of a number of anticancer agents against CRC.</jats:p>
<jats:p>Cell lines were generated by enzymatic disaggregation method. The tissues were obtained after informed consent from patient. The tumor tissues were sectioned into to small pieces (∼1mm) and were digested using 0.5% Collagenase for 2-4 hrs at 37°C. The single cell suspensions were cultured in DMEM supplemented with 10% fetal bovine serum. The epithelial cells were propagated up to passage 2 and were cryopreserved for further applications. Mutational (K-RAS, BRAF, PIK3CA, PTEN) and standard CRC specific marker (such as mucin) analyses were done in order to confirm the authenticity of these lines. The cell viability was evaluated using MTT assay following treatment with Standard of Care (SOC) agents (both single and combinations). Additionally for screening of selected FDA approved NSAIDs like Aspirin, Etoricoxib, Ibuprofen, Indomethacin, Mefenamic acid, Methotrexate and Nimesulide (repurposing) were performed at their respective CMAX concentrations after 48 hrs of treatment.</jats:p>
<jats:p>Our results showed that, the SOCs when tested as single agents did not show any significant cell death. However, when treated in combination with Irinotecan, Fluorouracil and Leucovorin in the presence and absence of Avastin a significant augmentation of cell death was observed in both the cell lines. Further, Mefenamic acid, Methotrexate, Ibuprofen and Aspirin exhibited a significant cytotoxic effect in both cell lines after 48 hrs of treatment. Together, these results demonstrate that COX-2 inhibitors could be developed as a as an effective anticancer therapeutics for colorectal cancer treatment and primary cell line model provides a rapid and more reliable screening platform.</jats:p>
<jats:p>Citation Format: Rohini Nair, Aninda Basu, Muthu Dhandapani, Biswanath Majumder, Baraneedharan Ulaganathan. Generation of primary cell lines from advanced colon cancer: a tool for screening novel anticancer molecules. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2147. doi:10.1158/1538-7445.AM2013-2147</jats:p>