KLF6-SV1 overexpression accelerates human and mouse prostate cancer progression and metastasis
J. Clin. Invest. Goutham Narla, et al. 118:2711 doi:10.1172/JCI34780 [
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Figure 6Targeted KLF6-SV1 inhibition dramatically increases spontaneous apoptosis. (
A) qRT-PCR and Western blot analysis of the PC3M cell line transfected with si-NTC and si-SV1 demonstrated significant downregulation of KLF6-SV1 mRNA and protein at 72 and 96 h. (
B) Fluorescence-activated cell sorting analysis of si-NTC– and si-SV1–transfected PC3 cells at 72 and 96 h. On average, si-SV1 treatment increased cell death 5-fold. Data represent the average of 3 independent experiments. Numbers within plots indicate percent hypodiploid (i.e., apoptotic) cells. (
C) Western blot analysis demonstrated marked upregulation of caspase-3 and -8 and increased poly(ADP-ribose) polymerase (PARP) cleavage in si-SV1–transfected cell lines. (
D) qRT-PCR and Western blot analysis of si-NTC– and si-SV1–transfected cells for NOXA and Bcl-2 demonstrated marked upregulation of the proapoptotic NOXA with concomitant downregulation of Bcl-2. Numbers above actin blots in
A and
C represent densitometric analysis of the protein bands, expressed as fold change relative to control and normalized to actin. In
A,
B, and
D, lanes were run on the same gel but were noncontiguous (black lines). Each Western blot shown is representative of 3 independent experiments. ***
P < 0.0001 versus control.
