To explore the correlation between multi-drug resistance-associated protein 4(MRP4) and the sensitivity of rectal cancer to radiation.A total of 95 patients with advanced rectal cancer and received radiation therapy between January 2000 and January 2009. MRP4 and P53 protein expression in the paraffin-embedded specimen were detected by immunohistochemistry. Logistic regression analysis was used to evaluate factors associated with the sensitivity of rectal cancer to radiation.Forty patients(42%) were sensitive to radiation therapy, of whom 10(11%) achieved pathological complete remission. Fifty-five patients were (58%) not responsive to radiation. Patients with low expression of MRP4 had a 66.7%(24/36) response rate, significantly higher than that of patients with high MRP4 expression (29.1%,16/59)(P<0.05). Patients with low expression of P53 had a 63.9%(23/36) response rate, significantly higher than that of patients with high P53 expression(28.8%,17/59)(P<0.01). The response rate after long course radiation therapy was 83.3%(20/24), significantly higher than that of patients who underwent short and medium course radiation[(31.3%, 5/16) and(27.3%,15/55)](P<0.01). Multivariate Logistic regression analysis showed radiation regimen, the expression of P53 and MRP4 protein were independently associated with the sensitivity of rectal cancer to radiation(P<0.05).MRP4 may serve as a predictive marker for the sensitivity of rectal cancer to preoperative radiation.