ZNF652-Induced circRHOT1 Promotes SMAD5 Expression to Modulate Tumorigenic Properties and Nature Killer Cell-Mediated Toxicity in Bladder Cancer via Targeting miR-3666

Figure 2

circRHOT1 knockdown attenuates BC cell proliferation, migration, and susceptibility to NK cells. (a) HT-1376 and HTB9 cells were stably transfected with si-NC, si-circRHOT1#1, and si-circRHOT1#2. Expression level of circRHOT1 in transfected cells was detected by qRT-PCR. (b, c) Proliferation of transfected HT-1376 (b) and HTB9 (c) cells determined by CCK-8 assay. (d, e) Measurement of proliferation of HT-1376 and HTB3 cells (d) with downregulated circRHOT1 by EDU assay, and analysis results (e). (f, g) Measurement of migration of HT-1376 or HTB3 cells (f) with downregulated circRHOT1 by Transwell migration assay, and statistical analysis results (g). (h) Killing of HT-1376 or HTB3 cells by circRHOT1 knockdown measured by 51Cr release assay. (i) Numbers of NK cells and circRHOT1-knockdown cells measured by perforin polarization assay. (j, k) Conjugation between NK cells and HT-1376 (j) or HTB9 cells (k) with downregulated circRHOT1 assessed by conjugation assay. Experiments were conducted in triplicate. , , and .

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)