HSCC tissues, adjacent normal tissues and lymph node metastases from 5 HSCC patients were collected and stored at -20℃.Fresh tumor tissue was stored in GEXSCOPE® tissue preservation solution (Singleron) and shipped on ice to the Singleron laboratory as soon as possible. Single cell suspension was loaded onto a microfluidic chip and single cell RNA-seq libraries were constructed according to the manufacturer's instructions (Singleron GEXSCOPE Single Cell RNAseq Library Kit, Singleron Biotechnologies).