Background
Methods
Cell lines and clinical tissues
Quantitative real-time polymerase chain reaction analysis, western blotting analysis, and immunofluorescence assays
Transfection experiment
Immunohistochemistry
In vivo circRNA precipitation
Knockdown or overexpression of circTRIM33–12
Cell proliferation, migration, and Matrigel invasion assay
Flow cytometry assay of the cell cycle
RNA immunoprecipitation (RIP)
Luciferase reporter assay
In vivo tumor growth and metastasis assays
Statistical analysis
Results
Dysregulated TRIM33-derived circTRIM33–12 in HCC
Variable | circTRIM33–12 | P value | |
---|---|---|---|
Low | High | ||
Age (years) | |||
≤ 50 | 44 | 46 | 1.000 |
> 50 | 56 | 55 | |
Sex | |||
Female | 13 | 17 | 0.553 |
Male | 87 | 83 | |
HBsAg | |||
Negative | 21 | 20 | 1.000 |
Positive | 79 | 80 | |
HCVAb | |||
Negative | 3 | 4 | 1.000 |
Positive | 97 | 96 | |
Liver cirrhosis | |||
No | 87 | 83 | 0.553 |
Yes | 13 | 17 | |
Serum AFP, ng/mL | |||
≤ 20 | 30 | 47 | 0.02 |
> 20 | 70 | 53 | |
Serum ALT, U/L | |||
≤ 75 | 86 | 86 | 1.000 |
> 75 | 14 | 14 | |
Tumor size (diameter, cm) | |||
≤ 5 | 49 | 66 | 0.022 |
> 5 | 51 | 34 | |
Encapsulation invasion | |||
Absent | 38 | 58 | 0.007 |
Present | 62 | 42 | |
Vascular invasion | |||
No | 51 | 73 | 0.002 |
Yes | 49 | 27 | |
Tumor number | |||
Single | 75 | 88 | 0.028 |
Multiple | 25 | 12 | |
TNM | |||
I/ II | 73 | 71 | 0.875 |
III/IV | 27 | 29 |
OS | ||||
---|---|---|---|---|
Multivariate | ||||
Factors | Univariate, P | HR | 95% CI | P value |
Sex (female vs. male) | 0.106 | NA | ||
Age (years) (≤50 vs. > 50) | 0.118 | NA | ||
HBsAg (positive vs. negative) | 0.730 | NA | ||
HCVAb (positive vs. negative) | 0.860 | NA | ||
Liver cirrhosis (yes vs. no) | 0.598 | NA | ||
Serum AFP, ng/mL (≤20 vs. > 20) | 0.424 | NA | ||
Serum ALT, U/L (≤75 vs. > 75) | 0.627 | NA | ||
Tumor size (diameter, cm) (> 5 vs ≤ 5) | 0.004 | 1.172 | 1.066–2.750 | 0.026 |
Tumor number (multiple vs. single) | 0.175 | NA | ||
TNM (III/IV vs I/II.) | 0.121 | NA | ||
CircTRIM33–12 expression (high vs. low) | 0.001 | 0.504 | 0.932–1.942 | 0.007 |
Cumulative Recurrence | ||||
---|---|---|---|---|
Multivariate | ||||
Factors | Univariate, P | HR | 95% CI | P value |
Sex (female vs. male) | 0.161 | NA | ||
Age (years) (≤50 vs. > 50) | 0.184 | NA | ||
HBsAg (positive vs. negative) | 0.419 | NA | ||
HCVAb (positive vs. negative) | 0.708 | NA | ||
Liver cirrhosis (yes vs. no) | 0.489 | NA | ||
Serum AFP, ng/mL (≤20 vs. > 20) | 0.182 | NA | ||
Serum ALT, U/L (≤75 vs. > 75) | 0.622 | NA | ||
Tumor size (diameter, cm) (> 5 vs ≤ 5) | < 0.001 | 2.055 | 1.335–3.163 | 0.001 |
Tumor number (multiple vs. single) | 0.131 | NA | ||
TNM (III/IV vs I/II.) | 0.099 | NA | ||
CircTRIM33–12 expression (high vs. low) | 0.001 | 0.534 | 0.344–0.830 | 0.005 |