Figures & data
Table 1 Correlation Between Circ_0062020 Expression and Clinical Clinicopathological Parameters of PCa
Figure 1 Radiation treatment lead to downregulate circ_0062020 expression in PCa cells. (A) The expression of circ_0062020 in radiosensitive (n=30), radioresistant (n=30) or normal (n=30) PCa tissues was analyzed by qRT-PCR. (B) Circ_0062020 expression was determined by qRT-PCR in PCa and normal cells. (C, D) The effect of radiation treatment on circ_0062020 expression was determined by qRT-PCR in PCa cells. *P<0.05.
Figure 2 Circ_0062020 knockdown aggravated IR-induced suppression in proliferation, metastasis and promotion in apoptosis. (A) Circ_0062020 expression was tested by qRT-PCR in PCa cells transfected with si-circ_0062020 or si-NC. (B, C) Survival fraction of PCa cells transfected with si-circ_0062020 or si-NC and treated by different doses of IR was determined by colony formation assay. (D–H) PCa cells were treated with IR before follow-up experiments. (D) Effect of si-circ_0062020 on the viability of IR-treated PCa cells was determined by CCK8 assay. (E, F) The migration and invasion were monitored by transwell assay in IR-treated PCa cells transfected with si-circ_0062020 or si-NC. (G) Wound healing assay was performed to assess the migration of IR-treated PCa cells transfected with si-circ_0062020 or si-NC. (H) Flow cytometry was used to evaluate the effect of si-circ_0062020 on apoptosis in IR-treated PCa cells. *P<0.05.
Figure 3 Circ_0062020 interacted with miR-615-5p. (A) The sequences contains wide type or mutant binding sites of circ_0062020 in miR-615-5p was predicted by CircRNA interactome online tool. (B) MiR-615-5p expression level was detected by qRT-PCR in PCa cells transfected with miR-615-5p mimic or miRNA NC. (C, D) Dual-luciferase reporter system was devoted to detecting the potential interaction between circ_0062020 and miR-615-5p in PCa cells. (E, F) MiR-615-5p expression level was detected by qRT-PCR in radiosensitive, radioresistant or normal PCa tissues and PCa cells or normal cells. (G, H) The effect of radiation treatment on miR-615-5p expression was detected by qRT-PCR in PCa cells. *P<0.05.
Figure 4 MiR-615-5p inhibitor reversed si-circ_0062020-induced suppression in proliferation, metastasis and promotion in apoptosis in IR-treated PCa cells. (A) The effect of si-circ_0062020 and miR-615-5p inhibitor on miR-615-5p expression was detected by qRT-PCR in IR-treated PCa cells. (B) The effect of si-circ_0062020 and miR-615-5p inhibitor on viability was tested by CCK8 assay in IR-treated PCa cells. (C, D) The migration and invasion were monitored by transwell assay in IR-treated PCa cells transfected with si-circ_0062020, si-circ_0062020 + miR-615-5p or negative control. (E) Wound healing assay was performed to assess the metastasis of IR-treated PCa cells transfected with si-circ_0062020, si-circ_0062020 + miR-615-5p or negative control. (F) Flow cytometry was used to evaluate the effect of si-circ_0062020 and miR-615-5p inhibitor on apoptosis in IR-treated PCa cells. *P<0.05.
Figure 5 MiR-615-5p bound to TRIP13 3ʹUTR in PCa cells. (A) The potential binding sites between TRIP13 3ʹ UTR and miR-615-5p was presented by TargetScan online database. (B, C) The luciferase activity was detected by dual-luciferase reporter system in PCa cells transfected with miR-615-5p mimic + WT-TRIP13-3ʹUTR or negative controls. (D) QRT-PCR was used to detect the expression of TRIP13 mRNA expression in radiosensitive, radioresistant PCa tissues or normal tissues. (E) The protein expression level of TRIP13 was detected by Western blot in PCa cells or normal cells. (F, G) The effect of IR treatment on TRIP13 mRNA expression was presented by qRT-PCR in PCa cells. *P<0.05.
Figure 6 TRIP13 overexpression allayed miR-615-5p mimic-mediated repression in TRIP13 protein expression, proliferation, metastasis and stimulation in apoptosis in IR-treated PCa cells. (A) The transfection efficiency of pc-TRIP13 was determined by Western blot. (B) Western blot was applied to detect TRIP13 protein expression in IR-treated PCa cells transfected with miR-615-5p mimic, miR-615-5p mimic+pc-TRIP13 or negative controls. (C) The effect of miR-615-5p mimic and pc-TRIP13 on viability was presented by CCK8 assay in IR-treated PCa cells. (D, E) Transwell assay was used to assess the migration and invasion in IR-treated PCa cells transfected with miR-615-5p mimic, miR-615-5p mimic + pc-TRIP13 or negative controls. (F) Wound healing assay was performed to estimate the metastasis in IR-treated PCa cells transfected with miR-615-5p mimic, miR-615-5p mimic + pc-TRIP13 or negative controls. (G) The apoptosis rate was tested by flow cytometry assay in IR-treated PCa cells transfected with miR-615-5p mimic, miR-615-5p mimic + pc-TRIP13 or negative controls. *P<0.05.
Figure 7 MiR-615-5p inhibition relieved si-circ_0062020-induced restraint in TRIP13 protein expression in IR-treated PCa cells. (A) Western blot was used to examine TRIP13 protein expression in IR-treated PCa cells transfected with si-circ_0062020 + miR-615-5p inhibitor or negative controls. *P<0.05.
Figure 8 Circ-0062020 knockdown inhibited PCa tumor growth in vivo. (A, B) Tumor volume and tumor weight of nude mice infected with PCa cells that transfected with sh-circ_0062020 or sh-NC were measured. (C, D) The expression of circ-0062020 and miR-615-5p was performed by qRT-PCR in xenograft PCa tumors. (E) Western blot was performed to assess TRIP13 protein expression in xenograft PCa tumors. *P<0.05.
