Method for adjusting in-vitro migration of prostatic cancer cells through miR-203/SNAI2 axis
A mir-203, psin-mir-203 technology, applied in the field of molecular detection, can solve the problem that the interaction between miRNAs and SNAI2 has not been well elucidated
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[0009] 1. Cell Culture
[0010] Five prostate cancer cell lines (DU145, PC3, 22Rv1, LNCaP, and C4-2) and two immortalized and untransformed prostate epithelial cell lines (PZ-HPV-7 and RWPE1) were obtained from ATCC, USA. Cells were cultured according to the standard manual of ATCC. Human umbilical vein endothelial cells (HuVECs) were purchased from Life Technologies (Carlsbad, California, USA) and cultured according to the instructions.
[0011] 2. Semi-quantitative RT-PCR and qRT-PCR
[0012] Semi-quantitative RT-PCR and qRT-PCR were performed as previously described. The primer sequences are as follows: 5´-GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCTAGTG-3´ (specific RT primer formiR-203), 5´-GTGCAGGGTCCGAGGT-3´ (real time PCR, miR-203, forward), 5´-GCCGCGTGAAATGTTTAGG-3´ (real time PCR , miR-203, reverse); 5´-CTCGCTTCGGCAGCACA-3´ (real time PCR, U6, forward), 5´-AACGCTTCACGAATTTGCGT-3´ (real time PCR, U6, reverse).
[0013] 3. Transwell infection detection
[0014] ...
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