Figure 5.

LDLR KD mimics miR-92b, while LDLR overexpression mitigates the influence of miR-92b, in human cervical carcinoma cells. (a, b) The proliferation of SiHa and CaSki cells with transient transfection of LDLR siRNAs was greater compared to cells transfected with scrambled vector as assessed by a CCK-8 assay. (c, d) siRNA-mediated KD of LDLR elicited G1 arrest in (c) CaSki and (d) SiHa cells incubated with nocodazole. (e, f) SiHa and CaSki cells with transient transfection of LDLR-targeted siRNAs exhibited decreased apoptosis as determined by incubation with 50 μg/ml PI and FITC-conjugated Annexin V, and evaluated on a flow cytometer. (g–j) The proliferation of SiHa and CaSki cells with transient transfection of LDLR-targeted siRNAs was quantified by CCK-8 after a 48-hour incubation with paclitaxel (0–160 nM) and cisplatin (0–40 μM) to assess chemosensitivity. Transiently transfected si-LDLR-1 and si-LDLR-2 attenuated the chemosensitivity of SiHa and CaSki cells to paclitaxel and cisplatin relative to NCs. (k, l) The proliferation of SiHa and CaSki cells with transient transfection of miR-92b mimics with LDLR-WT, LDLR-mut, or NC was assessed by CCK-8 assay. (m, n) The cell-cycle progression of SiHa and CaSki cells with transient transfection of miR-92b mimics with LDLR-WT, LDLR-mut, or NC was determined by flow cytometry upon incubation with nocodazole. (o, p) Apoptosis of SiHa and CaSki cells with transient transfection of miR-92b mimics with LDLR-WT, LDLR-mut, or NC was determined by incubation with 50 μg/ml PI and FITC-conjugated Annexin V, and evaluated on a flow cytometer.

Data are represented as means ± SEMs. *p < 0.05, **p < 0.01.

CCK-8, Cell Counting Kit-8; FITC, fluorescein isothiocyanate; KD, knockdown; LDLR, low-density lipoprotein receptor; NC, negative control; PI, propidium iodide; SEM, standard error of the mean; siRNA, short interfering RNA; WT, wild type.