Tumor tissues were dissociated using 2mg/ml Collagenase II (Sigma, C2-28-100MG) for 1h in 37℃ water bath. Digestion was terminated by wash with 5ml DMEM twice. Cells were passed through 40μm cell strainer to get single cell suspension. Cell viability were confirmed using AO/PI reagent (Absin, abs9727-100T). Then, single cell suspension was washed and resuspended in DPBS containing 0.02%BSA for single cell RNA-seq library construction. Construction of single cell RNA-seq library was performed using 10x Genomics Chromiun Single-Cell 3' kit(V3) according to the manufacturer's instructions. Briefly, cells were loaded onto the 10x Genomics single-cell-A chip for a target capture of 8000 cells. After droplet generation, sample were transferred into a PCR tube and performed reverse transcription and library construction.