Effect Of Dihydroartemisinin On The MRP2 Expression Of Human Ovarian Cancer Cell Line SKOV3/DDP In Nude Mice

Objective:To investigate the effects of different doses of dihodryartemisinin(DHA) on the human ovarian cancer cell line SKOV3/DDP in nude mice and the expression of MRP2. Methods:Culturing the ovarian cancer cell line SKOV3/DDP to the logarithmic growth phase, and inoculate the cells into the nude mice armpit subcutaneous tissue to establish transplanted tumor model.The tumor bearing mice were randomly divided into 4 groups, respectively as the blank control group(sesame oil),DDP group(2mg / kg),DDP(2mg / kg) + low dose group of DHA(100mg / kg) and DDP(2mg / kg) + high dose group of DHA(200mg / kg), each group included five nude mice. Cisplatin by intraperitoneal injection two days one time, a total of 5 times; DHA by orally administration, once a day, a total of 10 days. The nude mice were sacrificed in eleventh day and weighed the tumor to calculate the inhibition rate of each intervention group. The expression of MRP2 protein was detected by immunohistochemistry and the expression of m RNA MRP2 was detected by real-time RT-PCR method. Results:(1)The average tumor weight(g) of blank group, DDP group, DDP+ low /high doses group of DHA were 1.10, 1.05, 0.79, 0.53, and the inhibition rate in intervention groups respectively 4.19%, 28.42%, 51.91%; Compared with DDP group and blank group, there was no statistical significance(P>0.05);each of the rest two groups, the difference was statistics significant(P<0.05).(2)HE staining: the tumor cells in blank group and DDP group were arranged in order, pathological karyokinesis phase is common, almost no nuclear pyknosis,fragmentation and no visible necrotic area;the tumor cells in DDP+low/high doses group of DHA were arranged in disorder, and have irregular shapes, we observed cytoplasm rarefaction,nuclear pyknosis hyperchromatic and fragmentation in these cells, the necrosis regions were filled with red cells.(3)Immunohistochemistry: MRP2 was mainly expressed in the cell membrane, the highest expression groups of MRP2 protein were blank group and DDP group,and there have no statistically significance in it(P>0.05); DDP+ low/high doses group of DHA were low expressed, and obviously lower than the blank group(P<0.05).(4)real-time RT-PCR: The MRP2 m RNA expression in DDP+ low/high doses group of DHA was significantly decreased, compared with the control group, the difference was statistically significant(P<0.05). Conclusion:DHA may enhance the sensitivity of cells to cisplatin by inducing the low expression of MRP2, reverse the cisplatin resistance in nude mice, and the intensity of the action was positively correlated with a certain concentration range.