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. 2021 Feb 26;9:644397. doi: 10.3389/fcell.2021.644397

FIGURE 1.

FIGURE 1

circ_0057558 knockdown suppressed prostate cancer cell proliferation in vitro and in vivo. (A,B) Lentivirus expressing specific shRNAs targeting circ_0057558 (shcirc#1 and shcir#2) and control shRNAs (shNC#1 and shNC#2) were infected into 22RV1 (A) and DU145 cells (B). circ_0057558 expression was detected by qRT-PCR at 48 h post infection. Wild-type cells (WT) that had no treatment were used as negative control. (C,D) CCK-8 assay was carried out to detect proliferation in circ_0057558 knockdown group (shcirc#1) and control group (shNC#1). (E,F) Colony formation assay was conducted to determine the colony-forming ability of circ_0057558 knockdown group (shcirc#1) and control group (shNC#1). (G,H) Cell cycle analysis detected by PI staining and flow cytometry analysis. (I) CyclinD1 and CyclinB1 were detected by western blotting. (J–M) 22RV1 cells infected with circ_0057558 shRNA (shcirc#1) or control shRNA (shNC#1) were transplanted into nude mice (n = 6 per group). The tumor growth curves (J), as well as the photos and weight (K) of xenografts on 33 days after inoculation are shown. Immunofluoresence staining with anti-Ki-67 (L) was carried out to assess cell proliferation in xenografts. Scale bar: 50 μm. (M) The Kaplan–Meier plot of survival duration in nude mice transplanted with 22RV1 inoculation infected with shcirc#1 or shNC#1 (n = 12 per group). Data were expressed as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 versus shNC#1; #P < 0.05 versus shNC#2.