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2017 ; 14
(3
): 3371-3378
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Downregulation of PDCD4 by miR-21 suppresses tumor transformation and
proliferation in a nude mouse renal cancer model
#MMPMID28927090
Yuan H
; Xin S
; Huang Y
; Bao Y
; Jiang H
; Zhou L
; Ren X
; Li L
; Wang Q
; Zhang J
Oncol Lett
2017[Sep]; 14
(3
): 3371-3378
PMID28927090
show ga
Programmed cell death 4 (PDCD4) is known to suppress neoplastic transformation,
cell proliferation and metastasis, and to be downregulated by microRNA-21
(miR-21) in renal cell carcinoma (RCC) cell lines and tissues. The aim of the
present study was to investigate the roles of and association between PDCD4 and
miR-21 in a nude mouse renal cancer model. A total of 24 BALB/c male nude mice
were randomly assigned into the following three groups: Negative control (NC;
n=8), miR-21 inhibitor (n=8) and miR-21 mimic (n=8). Subsequently, renal cell
adenocarcinoma 786-O cells were subcutaneously transplanted into the armpits of
the mice, which were then injected daily with NC small interfering (si)RNA,
precursor-miR-21 (mimic) or anti-miR-21 (inhibitor). Tumors were removed from the
mice and weighed 16 days following 786-O cell transplantation. In addition, the
expression of miR-21 and PDCD4 mRNA in cancer tissues was analyzed using reverse
transcription-quantitative PCR. The expression of PDCD4 protein in cancer tissues
was also examined using immunohistochemistry and western blotting. Furthermore,
786-O cells were transfected with PDCD4 siRNA or NC siRNA, and the effects of
silencing PDCD4 on tumor cell growth, proliferation and invasion were
investigated using soft agar colony formation, EdU cell proliferation assay and
Transwell migration and invasion assays. Another 16 BALB/c male nude mice were
randomly assigned into two groups as follows: NC (n=8) and PDCD4 siRNA (n=8). The
786-O cells were subcutaneously transplanted into the armpits of the mice, which
were subsequently injected daily with NC siRNA or PDCD4 siRNA. The tumors were
removed and weighed 16 days following transplantation. Compared with the NC
group, tumor weight in the miR-21 mimic group was significantly increased. By
contrast, tumor weight in the miR-21 inhibitor group was significantly decreased.
Similar to the results observed in human renal cancer tissue and cell lines,
miR-21 expression in the nude mouse renal cancer models was significantly
upregulated in the miR-21 mimic group compared with the NC group, while it was
significantly lower in the miR-21 inhibitor group. Furthermore, there was a
significant reduction in PDCD4 protein levels in the miR-21 mimic group and a
significant increase in the miR-21 inhibitor group compared with the NC, whereas
PDCD4 mRNA expression was not significantly altered. In the EdU proliferation
assay, the mean percentage of new cells that incorporated EdU was 28.6% in the NC
siRNA group and significantly increased to 44.7% in PDCD4 siRNA transfected
cells. In the soft agar colony formation assay, Transwell and migration and
invasion assays, a significant increase in colony formation, migration and
invasion capacity in PDCD4 siRNA-transfected cells was observed compared with the
NC. Furthermore, compared with the NC group, tumor weight in the PDCD4 siRNA
group was significantly increased. Similar to the results observed in human renal
cancer tissue and cell lines, miR-21 promoted cancer cell hyperplasia and
proliferation, and post-transcriptionally downregulated PDCD4 protein expression,
in the nude mouse renal cancer model. The results of the present study and
previous studies indicate that PDCD4 and miR-21 serve an important role in renal
cancer. Thus, increasing PDCD4 expression or inhibiting miR-21 expression may
constitute effective novel therapeutic strategies for the treatment of renal
cancer.
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