After transfec tion with miR 370 mimics or inhibitor, the expression of miR 370 was overexpressed and downregulated, respect ively, Too, the mRNA and protein levels of FoxM1 have been inhibited with miR 370 mimics and elevated with miR 370 inhibitor, so the expression of miR 370 was negatively associated to that of FoxM1 in K562 cells, Meanwhile, the expres sion of FoxM1 was further inhibited with HHT miR 370 mimics as compared with miR 370 mimics alone, The protein expression of FoxM1 was inhibited with HHT miR 370 mimics as compared with HHT NC and miR 370 mimics alone, Thus, FoxM1 had a function in improved sensitivity to HHT with upregulation of miR 370. To additional identify the purpose of FoxM1, we checked its perform in cell apoptosis induced by miR 370.
Following transfection with miR 370 mimics or FoxM1 siRNA, the two miR 370 mimics and FoxM1 siRNA induced cell apoptosis, Otherwise, miR 370 inhibitor and FoxM1 overexpression plasmid inhibited cell apoptosis, When FoxM1 overexpression plasmid inhibitor compound libraries was transfected into K562 cells with miR 370 mimics to reverse the expression of FoxM1, the apoptosis was partially re versed. Yet, the FoxM1 siRNA that inhibited the miR 370 inhibitor induced overexpression of FoxM1 neutralized the inhibited apoptosis induced by anti miR 370 treatment. HHT mediated the upregulation of miR 370 in K562 cells To even further investigate the effect of HHT on miR 370 expression, we detected the expression of miR 370 and its target FoxM1 with incubation of HHT in K562 cells. In cells incubated with HHT at numerous concentrations for 72 h, the degree of mature miR 370 improved to about 37 fold and 77 fold that from the control, FoxM1 mRNA and pro tein expression was dose dependently downregulated, In addition, right after K562 cells have been incubated with HHT for 72 and 96 h, the level of miR 370 was upregulated, which was accompanied through the inhibition of FoxM1 mRNA and protein expression, Hence, HHT mediated the upregulation of miR 370 in K562 cells.
To even more detect the significance of miR 370 in HHT induced apoptosis, miR 370 inhibitor was inhibitor signaling inhibitor transfected into K562 cells with 0. 015 uM HHT for 72 h. The miR 370 inhibitor partially reversed HHT induced cell apoptosis as compared using the handle, MiR 370 expression was inhibited and FoxM1 expression was upregulated, There fore, the HHT miR 370 FoxM1 axis could possibly be a whole new regu latory mechanism in HHT induced apoptosis. Misregulation of miR 370 and FoxM1 in bone marrow from CML CP and CML BP patients MiR 370 expression was analyzed in bone marrow sam ples from 23 patients with newly diagnosed CML CP and ten with CML BP. The clinical qualities of CML patients are in Added file 4. Table S1. The degree of miR 370 was reduced in CML patients than healthful controls. In addition, miR 370 expression was reduce in CML BP than CML CP individuals, The mRNA level of FoxM1 was higher in patients with CML BP than CML CP, together with the expression lowest in healthy controls, which showed the negative association with miR 370 expression.