MiR-29c is downregulated in gastric carcinomas and regulates cell proliferation by targeting RCC2.
MiR-29c is downregulated in gastric carcinomas and regulates cell proliferation by targeting RCC2.
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miR-29c在胃癌中被下调，并通过靶向RCC2来调节细胞增殖。
DOI：
10.1186/1476-4598-12-15
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发表时间：
2013-02-25
期刊：
影响因子：
37.3
通讯作者：
Moriyama M
中科院分区：
文献类型：
--
作者：
Matsuo M;Nakada C;Tsukamoto Y;Noguchi T;Uchida T;Hijiya N;Matsuura K;Moriyama M
此前，利用miRNA微阵列，我们已经发现miR-29c在进展期胃癌中显著下调。在本研究中，我们研究了miR-29c在胃癌细胞中是否作为肿瘤抑制基因miRNA发挥作用。为此，我们验证了miR-29c在胃癌组织中的下调，并评价了miR-29c对胃癌细胞的生物学作用。在miR-29c转基因细胞中，细胞的增殖能力和软琼脂集落形成能力均显著降低。在未诱导细胞凋亡的情况下，miR-29c细胞BrdU掺入率和组蛋白H3(S10)磷酸化阳性细胞比例显著降低，提示miR-29c可能参与了细胞增殖的调控。为了解释miR-29c抑制细胞生长的机制，我们利用基因芯片技术研究了miR-29c与阴性对照细胞的差异表达基因(>；2倍)。在miR-29c转基因的MKN45、MKN7和MKN74细胞中，RCC2、PPIC和CDK6普遍下调，并且所有基因的3‘端非编码区都含有miR-29c靶序列。RCC2和PPIC在胃癌组织中表达上调，因此被认为是miR-29c在胃癌中的可能靶点。为了确定Rcc2和/或PPIC的下调是否参与了miR-29c对细胞生长的抑制，我们将针对Rcc2和PPIC的siRNAs导入MKN45，并测定了细胞存活率、BrdU掺入率和caspase活性。我们发现，Rcc2基因敲除降低了细胞存活率和BrdU掺入，但没有增加caspase活性，而PPIC基因敲除并没有增加caspase活性，这表明Rcc2基因的下调可能至少是miR-29c抑制细胞生长的部分原因。我们的发现表明miR-29c在胃癌细胞中可能具有肿瘤抑制作用，其表达降低可能通过Rcc2的异常表达而使肿瘤细胞具有生长优势。
Previously, using miRNA microarray, we have found that miR-29c is significantly downregulated in advanced gastric carcinoma. In the present study, we investigated whether miR-29c functions as a tumor-suppressor miRNA in gastric carcinoma cells. For this purpose, we verified the downregulation of miR-29c in gastric carcinoma tissues, and assessed the biological effect of miR-29c on gastric carcinoma cells. In miR-29c-transfected cells, both proliferation and colony formation ability on soft agar were significantly decreased. Although apoptosis was not induced, BrdU incorporation and the proportion of cells positive for phospho-histone H3 (S10) were significantly decreased in miR-29c-transfected cells, indicating that miR-29c may be involved in the regulation of cell proliferation. To explain the mechanism of growth suppression by miR-29c, we explored differentially expressed genes (>2-fold) in miR-29c-transfected cells in comparison with negative control transfected cells using microarray. RCC2, PPIC and CDK6 were commonly downregulated in miR-29c-transfected MKN45, MKN7 and MKN74 cells, and all of the genes harbored miR-29c target sequences in the 3’-UTR of their mRNA. RCC2 and PPIC were actually upregulated in gastric carcinoma tissues, and therefore both were identified as possible targets of miR-29c in gastric carcinoma. To ascertain whether downregulation of RCC2 and/or PPIC is involved in the growth suppression by miR-29c, we transfected siRNAs against RCC2 and PPIC into MKN45 and determined cell viability, the rate of BrdU incorporation, and caspase activity. We found that RCC2-knockdown decreased both cell viability and BrdU incorporation without any increase of caspase activity, while PPIC-knockdown did not, indicating that downregulation of RCC2 may be at least partly responsible for the growth suppression by miR-29c. Our findings indicate that miR-29c may have tumor-suppressive functions in gastric carcinoma cells, and that its decreased expression may confer a growth advantage on tumor cells via aberrant expression of RCC2.
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影响因子：
8
作者：
Saito, Y.;Suzuki, H.;Hibi, T.
通讯作者：
Hibi, T.
影响因子：
64.5
作者：
LIU, J;FARMER, JD;SCHREIBER, SL
通讯作者：
SCHREIBER, SL
影响因子：
3.7
作者：
Ribeiro-dos-Santos Â;Khayat AS;Silva A;Alencar DO;Lobato J;Luz L;Pinheiro DG;Varuzza L;Assumpção M;Assumpção P;Santos S;Zanette DL;Silva WA Jr;Burbano R;Darnet S
通讯作者：
Darnet S
DOI：
10.1073/pnas.90.24.11850
发表时间：
1993-12-15
影响因子：
11.1
作者：
KE, HM;ZHAO, YD;FRIEDMAN, J
通讯作者：
FRIEDMAN, J
影响因子：
1.6
作者：
Martineau-Thuillier, S;Andreassen, PR;Margolis, RL
通讯作者：
Margolis, RL