Introduction
Materials and methods
Study subjects
Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR)
Cell culture and grouping
Construction of overexpressed Gas5 plasmid, cell transfection, and gene detection
Western blot for the detection of Gas5 protein and proliferation and apoptosis related proteins [21‐23]
Cell counting kit-8 (CCK8) assay
Flow cytometry
Transwell assay
Statistical analysis
Results
Expression of Gas5 in CRC tissues and CRC cell lines
Correlation of Gas5 expression with clinicopathological features of patients with CRC
Clinicopathological features | N | Gas5 mRNA expression |
P
| Gas5 protein expression |
P
|
---|---|---|---|---|---|
Age (years) | |||||
≤ 60 | 79 | 0.59 ± 0.20 | 0.162 | 0.46 ± 0.14 | 0.691 |
> 60 | 47 | 0.54 ± 0.18 | 0.47 ± 0.13 | ||
Gender | |||||
Male | 74 | 0.56 ± 0.20 | 0.582 | 0.45 ± 0.11 | 0.214 |
Female | 52 | 0.58 ± 0.20 | 0.48 ± 0.16 | ||
Tumor size (cm) | |||||
≤ 5 | 45 | 0.64 ± 0.19 | 0.002 | 0.53 ± 0.15 | < 0.001 |
> 5 | 81 | 0.53 ± 0.19 | 0.42 ± 0.10 | ||
Invasive depth | |||||
T1 | 28 | 0.62 ± 0.22 | 0.185 | 0.47 ± 0.14 | 0.053 |
T2 | 44 | 0.59 ± 0.18 | 0.50 ± 0.15 | ||
T3 | 35 | 0.54 ± 0.21 | 0.44 ± 0.11 | ||
T4 | 19 | 0.50 ± 0.16 | 0.41 ± 0.10 | ||
Lymph node metastasis | |||||
With | 67 | 0.55 ± 0.17 | 0.153 | 0.45 ± 0.14 | 0.202 |
Without | 59 | 0.60 ± 0.22 | 0.48 ± 0.12 | ||
Distant metastasis | |||||
With | 80 | 0.55 ± 0.21 | 0.107 | 0.45 ± 0.12 | 0.103 |
Without | 46 | 0.61 ± 0.18 | 0.49 ± 0.15 | ||
Tumor differentiation | |||||
Well or moderate differentiation | 74 | 0.55 ± 0.21 | 0.158 | 0.47 ± 0.13 | 0.412 |
Poor or no differentiation | 52 | 0.60 ± 0.18 | 0.45 ± 0.14 | ||
TNM staging | |||||
I–II | 60 | 0.66 ± 0.19 | < 0.001 | 0.49 ± 0.12 | < 0.034 |
III–IV | 66 | 0.49 ± 0.17 | 0.44 ± 0.14 | ||
Tumor site | |||||
Right hemicolon | 68 | 0.58 ± 0.20 | 0.577 | 0.46 ± 0.15 | 0.684 |
Left hemicolon | 58 | 0.56 ± 0.20 | 0.47 ± 0.12 |
Correlation of Gas5 expression with the survival of CRC patients
Factors | Univariate analysis | Multivariate analysis | ||
---|---|---|---|---|
Chi square |
P
| HR (95% CI) |
P
| |
Tumor site | 2.24 | 0.135 | ||
Gender | 0.21 | 0.650 | ||
Age | 1.81 | 0.178 | ||
Tumor size | 1.53 | 0.216 | ||
Invasive depth | 5.64 | 0.303 | ||
Lymph node metastasis | 4.43 | 0.035 | 1.50 (0.93–2.43) | 0.099 |
Gas5 | 27.13 | < 0.001 | 0.52 (0.03–0.91) | 0.022 |
Distant metastasis | 20.14 | < 0.001 | 2.76 (1.54–4.93) | 0.001 |
Tumor differentiation | 5.80 | 0.016 | 2.00 (1.25–3.20) | 0.004 |
TNM staging | 34.67 | < 0.001 | 2.81 (1.56–5.04) | 0.001 |