Background
Methods
Cell culture
Cell fusion
Cell isolation and colony formation
Gene sequencing and differential gene expression analysis
Flow cytometry
Quantitative RT-PCR
Gene | Forward primer (5’-3’) | Reverse Primer (5’-3’) |
---|---|---|
AFP | AGACTGAAAACCCTCTTGAATGC | GTCCTCACTGAGTTGGCAACA |
CD133 | ACATGAAAAG ACCTGGGGG | GATCTGGTGTCCCAGCATG |
CD44 | TCCCAGACGAAGACAGTCCCTGGAT | CACTGGGGTGGAATGTGTCTTGGTC |
ALDH1A1 | CTGCTGGCGACAATGGAGT | CTGCTGGCGACAATGGAGT |
ABCB1 | GGGAGCTTAACACCCGACTTA | GCCAAAATCACAAGGGTTAGCTT |
EpCAM | AATCGTCAATGCCAGTGTACTT | TCTCATCGCAGTCAGGATCATAA |
Bcl-2 | GACTGAATCGGAGATGGAGACC | GCAGTTCAAACTCGTCGCCT |
β-actin | CATCCTCACCCTGAAGTACCC | AGCCTGGATAGCAACGTACATG |
Drug resistance
In vivo tumorigenicity assay
Animal care and ethics statement
Statistical analysis
Results
Gene expression of the fused cells is different from HepG2 cells
Gene ontology term | Cluster frequency | Genome frequency of use | Corrected p-value |
---|---|---|---|
Immune response | 10.2 % | 4.9 % | 2.74e-07 |
Immune system process | 12.4 % | 6.8 % | 3.77e-06 |
Response to virus | 2.6 % | 0.8 % | 0.00138 |
Regulation of apoptosis | 7.8 % | 4.3 % | 0.00508 |
Regulation of programmed cell death | 7.8 % | 4.4 % | 0.00840 |
Regulation of cell death | 7.8 % | 4.4 % | 0.00888 |
Fused cells exhibited increased drug resistance
Fused cells are highly tumorigenic
Cell type | Cell numbers injected | Tumor incidencea
| Latency (days)b
|
---|---|---|---|
HepG2 | 5 × 104
| 0/4 | — |
1 × 105
| 0/4 | — | |
1 × 106
| 1/2 | 14 | |
Fused cells | 5 × 104
| 2/4 | 14 |
1 × 105
| 3/4 | 14 | |
1 × 106
| 4/4 | 12 |