Fig 1: PIM1 overexpression compensated the effect of miR-3135b in CRC cells. (A) In HCT116 cells, transfection with pcDNA3-PIM1 increased PIM1 expression when compared with pcDNA3. pcDNA3-PIM1 also reversed the miR-3135b mimic-induced downregulation of PIM1. In HCT116 cells, PIM1 overexpression (B) promoted cell proliferation and compensated the miR-3135b overexpression-induced cell viability reduction; (C) upregulated 5-FU sensitivity; and (D) downregulated cell apoptosis. &&&P<0.001 vs. pcDNA3. **P<0.01, ***P<0.001 vs. pcDNA3. #P<0.05 and ###P<0.001 vs. miR-3135b mimic. PIM1, proviral integration site for Moloney murine leukemia virus 1; miR, microRNA; CRC, colorectal cancer; 5-FU, 5-fluorouracil.
Fig 2: miR-3135b targeted the PIM1 3'UTR. (A) FL, Trun1 and Trun2 of PIM1 3'UTR are presented. (B) Compared with miR-NC, the relative luciferase activity of HCT116 cells co-transfected with FL was significantly decreased by the miR-3135b mimic, whereas in the HCT116 cells co-transfected with miR-3135b mimic and Trun1 the decrease was significant, but the difference was less. The HCT116 cells co-transfected with Trun2 were not affected by the miR-3135b mimic. (C) The combining sites between miR-3135b and PIM1, as well as the Mut sequences of PIM1 are presented. (D) The miR-3135b mimic significantly inhibited the relative luciferase activity of HCT116 cells transfected with PIM1 3'UTR-WT, but not PIM1 3'UTR-Mut when compared with miR-NC. *P<0.05 and ***P<0.001 vs. miR-NC. miR, microRNA; PIM1, proviral integration site for Moloney murine leukemia virus 1; UTR, untranslated region; FL, full-length; Trun, truncated; Mut, mutant; WT, wild-type; NC, negative control.
Fig 3: PIM1 mRNA levels were inversely correlated with miR-3135b expression in CRC tumor samples. (A) PIM1 expression was higher in tumor tissue samples when compared with non-tumor tissue samples. (B) PIM1 expression was higher in the tumor recurrent group when compared with that of the tumor non-recurrent group. (C) An inverse correlation was observed between PIM1 mRNA levels and miR-3135b expression in CRC tumor tissue samples. ***P<0.001. PIM1, proviral integration site for Moloney murine leukemia virus 1; miR, microRNA; CRC, colorectal cancer.
Fig 4: miR-3135b inhibited PIM1 expression. (A) PIM1 was complementary to miR-3135b. The miR-3135b mimic significantly inhibited the (B) relative mRNA and (C) protein expression of PIM1 when compared with miR-NC. ***P<0.001 vs. miR-NC. miR, microRNA; PIM1, proviral integration site for Moloney murine leukemia virus 1; NC, negative control; UTR, untranslated region.
Supplier Page from Abcam for Anti-PIM1 antibody