Background
Methods
Cell culture and transfections
Patients and specimens
RNA extraction, cDNA synthesis, and real-time PCR assays
Primer | Sequence(5′ → 3′) |
---|---|
miR-99a-RT | GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCACAAGA |
miR-99a-forward | GCTGGAGAACCCGTAGATCCGAT |
miR-99a-reverse | GTGCAGGGTCCGAGGT |
miR-99a-probe | FAM-ATACGACCACAAGATCGG-MGB |
U6-RT | AAAATATGGAACGCTTCACGAATTTG |
U6-forward | CTCGCTTCGGCAGCACATATACT |
U6-reverse | ACGCTTCACGAATTTGCGTGTC |
U6-probe | FAM-CCATGCTAATCTTCTCTGTA-MGB |
Cell proliferation assay
Cell cycle analysis
Statistics
Results
miR-99a is down-regulated in bladder cancer cells
miR-99a is down-regulated in bladder cancer tissues compared with the corresponding adjacent non- neoplastic tissues
Low-level expression of miR-99a is associated with aggressive phenotypes of bladder cancer
Variables | Patients, n | |
---|---|---|
Total | Lower miR-99a | |
(n = 100) | (n = 53) | |
Histology
| ||
TCC | 83 | 35 |
TCC with aberrant differentiation | 17 | 18 |
Gender
| ||
Male | 75 | 38 |
Female | 25 | 15 |
Age
| ||
≥60 | 62 | 34 |
<60 | 38 | 19 |
Stage
| ||
Ta | 34 | 15 |
T1 | 25 | 10 |
T2 | 18 | 11 |
T3 | 13 | 9 |
T4 | 10 | 8 |
Grade
| ||
1 | 25 | 7 |
2 | 40 | 18 |
3 | 35 | 28 |
Smoker
| ||
Current | 30 | 15 |
Ex | 44 | 21 |
Never | 26 | 17 |
Recurrence
| ||
Yes | 47 | 21 |
No | 53 | 32 |
Progression
| ||
Yes | 33 | 21 |
No | 67 | 32 |