- 作者: Cheng-Der Wu, Yuan-Sung Kuo, Han-Chung Wu, Chin-Tarng Lin
- 作者服務機構: Institute of Pathology, College of Medicine, National Taiwan University , Taiwan, R.O.C.
- 中文摘要: --
- 英文摘要:
Background:
MiR-1 (microRNA-1) has been used as a positive control in some microRNA
experiments. We found that miR-1 transfection of nasopharyngeal carcinoma cells
reveals a typical apoptotic process as shown by time-lapse microscopy so we
investigated the mechanisms of miR-1 inducing apoptosis.
Methods:
To confirm that miR-1 induces apoptosis, we used Annexin V and TUNEL staining
and caspase assay. To determine that miR-1 directly targets genes that involve in
apoptosis, we analyzed microRNA and pathway databases, and cDNA expression
microarrays from miR-1 transfected cells. To demonstrate candidate miR-1 targeted
genes, we used qRT-PCR analysis and luciferase reporter vector assays. To assess the
miR-1 target gene PTMA (prothymosin alpha, ProTalpha) involves in apoptosis, we
used PTMA siRNA to knock down PTMA.
Results:
Annexin V and TUNEL staining and caspase assay confirm that miR-1 induces
nasopharyngeal carcinoma cell apoptosis. MiR-1 transfection of HeLa, Cal-27,
KYSE30 and NPC-TW06 cell lines which express low levels of endogenous miR-1
also induces apoptosis. However, miR-1 transfection of cell lines such as SW620,
HepG2, HEK-293T, SAS and PC-13 which express high levels of endogenous miR-1
does not result in apoptosis. MiR-1 directly targets PTMA gene. PTMA siRNA and
miR-1 accelerate the apoptotic process in cells treated with apoptosis inducers.
Conclusions:
The exogenous expression of miR-1 induces apoptosis in a number of cell lines. This
is a model of microRNA-induced cell apoptosis. The PTMA is one of miR-1 target
genes which involve in miR-1 inducing apoptosis. MiR-1 and PTMA siRNA may
have potential applications in cancer therapies. - 中文關鍵字: --
- 英文關鍵字: nasopharyngeal carcinoma (NPC); microRNA-1 (miR-1); PTMA (prothymosin alpha; ProTalpha); apoptosis