Background
Materials and methods
Clinical specimens collection
Parameters | Cases | circular RNA circ-CPA4 | P value | |
---|---|---|---|---|
High | Low | |||
Age | 0.721 | |||
≥ 55 | 35 | 20 | 15 | |
< 55 | 15 | 5 | 10 | |
Gender | ||||
Male | 25 | 13 | 12 | 0.832 |
Female | 25 | 12 | 13 | |
TNM stage | 0.023 | |||
I/II | 18 | 9 | 9 | |
III/IV | 32 | 16 | 16 | |
Pathological Type | 0.593 | |||
Suqamous | 21 | 10 | 11 | |
Adenocarcinoma | 12 | 7 | 5 | |
Large Cell Lung cancer | 17 | 8 | 9 | |
Smoking status | 0.098 | |||
Non-smoker | 14 | 7 | 7 | |
Smoker | 16 | 7 | 9 | |
Heavy smoker | 20 | 11 | 9 | |
Lymphatic Metastasis | 0.019 | |||
No | 29 | 13 | 16 | |
Yes | 21 | 12 | 9 |
Parameters | Cases | Let-7 miRNA | P value | |
---|---|---|---|---|
High | Low | |||
Age | 0.093 | |||
≥ 55 | 35 | 18 | 17 | |
< 55 | 15 | 8 | 7 | |
Gender | 0.482 | |||
Male | 25 | 13 | 12 | |
Female | 25 | 13 | 12 | |
TNM stage | 0.005 | |||
I/II | 18 | 14 | 4 | |
III/IV | 32 | 12 | 20 | |
Pathological Type | 0.983 | |||
Suqamous | 21 | 12 | 9 | |
Adenocarcinoma | 12 | 5 | 7 | |
Large Cell Lung cancer | 17 | 9 | 8 | |
Smoking status | 0.447 | |||
Non-smoker | 14 | 7 | 7 | |
Smoker | 16 | 6 | 10 | |
Heavy smoker | 20 | 13 | 7 | |
Lymphatic Metastasis | 0.042 | |||
No | 29 | 14 | 15 | |
Yes | 21 | 12 | 9 |
Parameters | Cases | PD-L1 mRNA | P value | |
---|---|---|---|---|
High | Low | |||
Age | 0.062 | |||
≥ 55 | 35 | 16 | 19 | |
< 55 | 15 | 5 | 10 | |
Gender | 0.523 | |||
Male | 25 | 14 | 11 | |
Female | 25 | 7 | 18 | |
TNM stage | 0.031 | |||
I/II | 18 | 12 | 6 | |
III/IV | 32 | 9 | 23 | |
Pathological Type | 0.511 | |||
Suqamous | 21 | 8 | 13 | |
Adenocarcinoma | 12 | 7 | 5 | |
Large Cell Lung cancer | 17 | 6 | 11 | |
Smoking status | 0.345 | |||
Non-smoker | 14 | 6 | 8 | |
Smoker | 16 | 7 | 9 | |
Heavy smoker | 20 | 8 | 12 | |
Lymphatic Metastasis | 0.011 | |||
No | 29 | 7 | 22 | |
Yes | 21 | 14 | 7 |
Cell culture and vectors transfection
Gene | Primer sequences (strand) |
---|---|
PD-L1 | Forward: 5′-GGAUAAGAACAUUAUUCAAdTdT-3′ |
Reverse: 5′-UUGAAUAAUGUUCUUAUCCdTdT-3′ | |
Si-circ-CPA4 | 5′-UUCUCCGAACGUGUCACGUTT-3′ |
Mimic | 5′-UGAGGUAGUAGGUUGUAUGGUU-3’ |
Inhibitor | 5′-AACCAUACAACCUACUACCUCA-3’ |
Real-time qPCR
Gene | Primer sequences (strand) |
---|---|
β-actin | Forward: 5′-CTCCATCCTGGCCTCGCTGT-3′ |
Reverse: 5′-GCTGCTACCTTCACCGTTCC-3′ | |
U6 | Forward: 5′-GACTATCATATGCTTACCGT-3′ |
Reverse: 5′-GGGCAGGAAGAGGGCCTAT-3′ | |
Circ-CPA4 | Forward: 5′- ACAGCATCTGGTGTGTGCTT-3′ |
Reverse: 5′-CCCTTTCCTGCAAAACTAGC-3′ | |
Let-7 | Forward: 5′-AGCAAGCTTTGGCACCCACCCGTAGAAC-3′ |
Reverse: 5′-TAAGGATCCGATGCAGGGACAAGGACAGAA-3′ | |
PD-L1 | Forward: 5′-GCCGAAGTCATCTGGACAAG-3’ |
Reverse: 5′-TCTCAGTGTGCTGGTCACAT-3’ | |
OCT4 | Forward: 5′-AGCGATCAAGCAGCGACTA-3’ |
Reverse: 5′-GGAAAGGGACCGAGGAGTA-3’ | |
SOX2 | Forward: 5′-CATCACCCACAGCAAATGAC-3’ |
Reverse: 5′-CAAAGCTCCTACCGTACCACT-3’ | |
Nanog | Forward: 5′-GCAGGCAACTCACTTTATCC −3’ |
Reverse: 5′-CCCACAAATCACAGGCATAG-3’ | |
ALDH1 | Forward: 5′-AGCCTTCACAGGATCAACAGA-3’ |
Reverse: 5′-GTCGGCATCAGCTAACACAA-3’ | |
IFN-γ | Forward: 5′-CCGAAACAGGCTCCCAACCG-3’ |
Reverse: 5′-GGGAAGACGCTCTCACCT-3’ | |
IL-4 | Forward: 5′-AACGGCTCGACAGGAACCT-3’ |
Reverse: 5′-ACTCTGGTTGGCTTCCTTCCA-3’ | |
IL-10 | Forward: 5′-ACAGCCGGGAAGACAATAAC-3’ |
Reverse: 5′- CAGCTGGTCCTTTGTTTGAAA-3’ |