Abstract: OBJECTIVE To investigate the relationship between miR-101 and sensitivity of cervical cancer stem cells to 5-fluorouracil. METHODS Flow cytometry was performed to isolate the cancer stem cells in Hela cell line. RT-qPCR analysis was performed to detect the expression of miR-101 in Hela cancer stem cells. MTT assay was performed to evaluate the viability of Hela cancer stem cells which were treated with miR-101 and 5-fluorouracil. Western blot analysis was performed to detect the expression of c-met, phosphorylation of PI3K and AKT, and activation of caspases in the Hela cancer stem cells. Flow cytometry assay was performed to measure the apoptotic rate of Hela cancer stem cells. RESULTS Expression of miR-101 in Hela cancer stem cells was significantly lower than that in the Hela non-stem cells. Anti-tumor effect of 5-fluorouracil could be significantly enhanced when the miR-101 was transfected into the Hela cancer stem cells. The results of western blot showed that the expression of c-met was down-regulated by miR-101, indicating that the c-met gene was the target of miR-101. Furthermore, the results of Western blot analysis showed that miR-101 inhibited the phosphorylation of PI3K and AKT by targeting c-met. As the results, the sensitivity of Hela cancer stem cells to the 5-fluorouracil-induced apoptosis was significantly enhanced, and the activation of caspase-9 and caspase-3 was promoted. CONCLUSION miR-101 increased the sensitivity of cervical cancer stem cells to 5-fluorouracil by down-regulating the expression of c-met.