Next Generation Sequencing from Tumor Tissues And Tumor-adjacent Tissues of Renal Cell Carcinoma

Tumor-adjacent tissues are very important for tumor research. This study aimed to identify hub genes in tumor-adjacent tissues of mice transplanted with 786-o renal cell carcinoma cells. 786-o cells were implanted in nude mice raised to 1 month of age, and normal tissues, tumor tissues, and adjacent tissues were obtained for RNA sequencing. All tissues were sequenced for transcriptome genes. This was the first systematic exploration of the hub genes in the tumor-adjacent tissues of mice transplanted with 786-o renal cell carcinoma cells, which provides a genetic basis for further in-depth study on the underlying regulatory mechanism of renal cell carcinoma. Ten 4-week-old male BALB/c nude mice were obtained from the Animal Center of Shanghai Tenth People’s Hospital (Shanghai, China), every five mice were kept in a cage, kept under specific pathogen-free conditions at 21±2°C and 55±10% humidity under a 12 h light/dark cycle, and fed normal chow with ad libitum access to water. All mice were treated humanely. The 786-o cells were cultured in a 10 cm dish (DMEM medium, 10% fetal bovine serum, PS double antibody). When the cell density was 70-80%, the cells were collected, mixed into a cell suspension with 100 µl sterile phosphate-buffered saline solution, and injected into the right back of a nude mouse at approximately the midpoint of the spinal column and right lower limb. Before injection, 0.5 cm of tissue was taken from the tail of each nude mouse. The mice were fed normally, and the growth of transplanted tumors was observed every day. Seven of the ten mice transplanted tumors successfully. One month later, three mice with tumor diameter of 1-1.5 cm were selected for the next study. Mice were euthanized in a carbon dioxide box, and the tumor and tumor-adjacent tissues (more than 1 cm from tumor tissue capsule) were removed. All tissues were stored in liquid nitrogen and sent to Shanghai Majorbio company for RNA-seq analysis. Different tissues of each mouse were self-compared, three mice were repeated three times.