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Rami James N. Aoun*, Sylvain Ferrandon, May Zin Hlaing, Joesph Fedro, Matthew Kalady
Department of Surgery, The Ohio State University, Columbus, OH

Improved oncological response of rectal cancer to neoadjuvant radiation therapy correlates with improved health outcomes. This response ranges from a complete response, or 0 on the AJCC-TRG scale, to little or no response, or a 3 on the scale. We previously demonstrated an association between increased SPATA20 expression and higher AJCC scores in a patient cohort. We hypothesize that knocking down SPATA20 in in-vitro rectal cell lines will increase their radiosensitivity.

HRT18 rectal cancer cell lines were transfected with a lentiviral construct containing small interfering RNAs (siRNA) targeting SPATA20 mRNA to create knockdown cell lines. Western blot was used for knockdown validation. Wildtype HRT18 and knockdown HRT18 cell lines were plated and irradiated with 10 Gy. Cell viability was assessed at the 48-hours interval using Annexin V/Propidium Iodide (PI) cell viability assay for both irradiated and non-irradiated controls.

Decreased expression was confirmed in knockdown HRT18 cell lines compared to wildtype HRT18 using Western blot. In the non-radiated controls, Annexin V/PI demonstrated no statistical difference (p=0.63) in cell death when comparing wildtype HRT18 (28.3%) to knockdown HRT18 cell lines (30.6%). In the radiated samples on the other hand, there was a statistically significant increase (p=0.015) in cell death when comparing wildtype HRT18 (33.9%) to knockdown cell lines (52.87%). Comparing non-radiated to radiated knockdown HRT18, there was also a statistically significant increase (p=0.022) in cell death (30.6% vs 52.87%).

Manipulation of SPATA20 correlates with radiation response, signifying it as a possible modulator of radiosensitivity in rectal cancer. Further investigation is warranted to validate this observation in animal models and explore potential pharmacological inhibitors.

Comparison of cell death between non-radiated and radiated samples of wildtype and knockdown HRT18 cell lines

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