Silencing of the CrkL gene reverses the doxorubicin resistance of K562/ADR cells through regulating PI3K/Akt/MRP1 signaling.

BACKGROUND: Doxorubicin is a first-line chemotherapy agent on human myelogenous leukemia clinical treatment, but the development of chemoresistance has largely limited curative effect. In this study, we aimed to evaluate the biological function and molecular mechanisms of CrkL to Doxorubicin resistance. METHODS: Quantitative reverse transcription-PCR (qRT-PCR) assay was performed to examine the expression of CrkL in K562 and K562/ADR cells. The expression of CrkL was silenced through RNA interference technology. MTT assay and flow cytometry were performed to detect the proliferation inhibition and apoptosis rate after CrkL siRNA transfection. The protein expression changes of PI3K/AKT/MRP1 pathway induced by CrkL siRNA were observed by Western Blot assay. Xenograft tumor model was carried out to observe tumor growth in vivo. RESULTS: We observed that silencing of CrkL could effectively increase apoptosis rate induced by doxorubicin and dramatically reversed doxorubicin resistance in K562/ADR cells. Further studies revealed knockdown CrkL expression suppressed PI3K/Akt/MRP1 signaling, which indicated CrkL siRNA reversed doxorubicin effect through regulating PI3K/Akt/MRP1 pathway. In addition, overexpression of MRP1 could evidently reduce apoptosis rate and reversed the inhibitory effects of doxorubicin resistance caused by CrkL siRNA on K562/ADR cells. Finally, in vivo experiments revealed that CrkL silencing acted a tumor-suppressing role in myelogenous leukemia via regulating PI3K/Akt/MRP1 signaling. CONCLUSION: Together, we indicated that CrkL is up-regulated in myelogenous leukemia cells and silencing of CrkL could reverse Doxorubicin resistance effectively. These results show a potential novel strategy for intervention chemoresistance in myelogenous leukemia during chemotherapy.

[Identification of ecological corridors for Tibetan antelope and assessment of their human disturbances in the alpine desert of Qinghai-Tibet Plateau].

The alpine desert of Qinghai-Tibet Plateau (QTP) provides the largest habitats for those endangered ungulates (e.g., Tibetan antelope, Tibetan Kiang and wild yak) on the earth. However, human disturbance especially infrastructure constructions (e.g., railway & highway) has increasingly fragmented the habitats of those endangered ungulates by disturbing and interrupting their ecological corridors for their seasonal migration. Aiming at identifying the potential ecological corridors for Tibetan antelope, a GIS-based model-Linkage Mapper was used to model and detect the potential ecological corridors of Tibetan antelope based on the principle of least cost path. Three categories of ecological corridors, i. e., closed (inside reserves), linking (linking the reserves) and open (starting from reserve but ending outside) corridors were distinguished by their spatial interactions with existing major national nature reserves (i.e., Altun, Kekexili and Qiangtang NNRs) in the alpine desert of QTP, and their spatial patterns, conservation status associated with human disturbance were also examined. Although our research indicated a general ecological integration of both habitats and ecological corridors in the alpine desert ecosystem, increasing human disturbance should not be ignored, which particularly partially undermined the functioning of those ecological corridors linking the nature reserves. Considering disadvantages of prevailing separate administrative structure of nature reserve on the effective conservation of ecological corridors for those endangered ungulates, a coordinative conservation network among these major national nature reserves should be established to ensure the unified trans-boundary conservation efforts and to enhance its overall conservation efficacy by sharing information, knowledge and optimizing conservation resources.