Research Paper Volume 13, Issue 2 pp 2604—2625

Knockdown of CENPF inhibits the progression of lung adenocarcinoma mediated by ERβ2/5 pathway

Knockdown of CENPF inhibits cell proliferation, migration, invasion and increases apoptosis of LUAD cells. (A) The protein level of CENPF in A549 and H1299 cell lines were higher than in normal cell lines BEAS-2B and other LUAD cells. GAPDH served as the internal control. (The corresponding gray value are shown in Supplementary Figure 3). (B) The knockdown efficiency of LV-CENPF sh or LV-NC transfected with A549 and H1299 cells was verified by RT-qPCR. *P C, D) MTT showed that CENPF knockdown suppressed the proliferative viability of cells in A549 and H1299 cells. *P E) Migration assays and invasion assays revealed that CENPF-KD decreased cell migration and invasion abilities of A549. (F, G) The related protein E-cadherin was significantly increased (P=0.009, Figure 4F; The corresponding gray value are shown in Supplementary Figure 3G), and N-cadherin and MMP2 were significantly decreased when compared with NC group (P=0.004, 0.012; The corresponding gray value are shown in Supplementary Figure 3H). (H) Quantified histograms of scratch experiment of A549 and H1299. (I) The cell percentage and DNA content were significantly increased in the G1 phase in the CENPF-KD group(P=0.011). (J) The expression of CCND1, CDK2 and CDK4 was significantly lowered in CENPF-KD group (P=0.022, 0.001, 0.002; The corresponding gray value are shown in Supplementary Figure 3M). (K) CENPF knockdown increased apoptosis of A549 and H1299 cell lines (P=0.001, 0.001). Each experiment was performed in triplicate and repeated three times. P values were calculated with two-tailed unpaired Student’s t test.

Figure 4. Knockdown of CENPF inhibits cell proliferation, migration, invasion and increases apoptosis of LUAD cells. (A) The protein level of CENPF in A549 and H1299 cell lines were higher than in normal cell lines BEAS-2B and other LUAD cells. GAPDH served as the internal control. (The corresponding gray value are shown in Supplementary Figure 3). (B) The knockdown efficiency of LV-CENPF sh or LV-NC transfected with A549 and H1299 cells was verified by RT-qPCR. *P < 0.05 vs CENPF-KD. (C, D) MTT showed that CENPF knockdown suppressed the proliferative viability of cells in A549 and H1299 cells. *P < 0.05 vs CENPF-KD. (E) Migration assays and invasion assays revealed that CENPF-KD decreased cell migration and invasion abilities of A549. (F, G) The related protein E-cadherin was significantly increased (P=0.009, Figure 4F; The corresponding gray value are shown in Supplementary Figure 3G), and N-cadherin and MMP2 were significantly decreased when compared with NC group (P=0.004, 0.012; The corresponding gray value are shown in Supplementary Figure 3H). (H) Quantified histograms of scratch experiment of A549 and H1299. (I) The cell percentage and DNA content were significantly increased in the G1 phase in the CENPF-KD group(P=0.011). (J) The expression of CCND1, CDK2 and CDK4 was significantly lowered in CENPF-KD group (P=0.022, 0.001, 0.002; The corresponding gray value are shown in Supplementary Figure 3M). (K) CENPF knockdown increased apoptosis of A549 and H1299 cell lines (P=0.001, 0.001). Each experiment was performed in triplicate and repeated three times. P values were calculated with two-tailed unpaired Student’s t test.