Study on the role of transient receptor potential C6 channels in esophageal squamous cell carcinoma radiosensitivity

Yi Zhang, Yongxing Zhang, Hong Fan, Yu Shi, Cheng Zhan, Qun Wang


Background: To study the effect of transient receptor potential C6 (TRPC6) channels on esophageal squamous cell carcinoma (ESCC) cell lines Eca109 cell cycle and to confirm whether TRPC6 channel is candidate radiosensitivity in vitro and in vivo.
Methods: We chose Eca109 cell line with a strong TRPC6 channels expression. Cell cycle was investigated after TRPC6 channel inhibitor SKF96365 treated with a 5 μM concentration. According to the results of cell cycle, radiation was performed. CCK-8 test was used to test the cell proliferation. Then we performed the same study in vivo. Total of 40 male nude mice were randomly divided into four groups as follows: SKF96365, radio, combined radio-SKF96365 and control. In SKF96365 group, 20 mg/kg 5 μM SKF96365 was injected into the abdominal cavity of the nude mice at day 5–11. In radiation group, the nude mice received radiotherapy 2 Gy per day at day 7–11. In combined radio-SKF96365 group, 20 mg/kg 5 μM SKF96365 was injected into the abdominal cavity of the nude mice at day 5–11 and 2 Gy radiotherapy was delivered to the tumor site at day 7–11. In control group, nude mice were injected saline into the abdominal cavity at day 5–11. General states of health were observed, the tumor size in volume was measured with calipers two times every week. Six weeks after seeding, mice were sacrificed by neck-break. The tumor size was measured in volume with caliper and in weigh with scale.
Results: Treatment with SKF96365 substantially increased the percentage of Eca109 cells in the G2/M phase and reduced that in G0/G1 phase in a time-dependent manner. Most of the cells (85.26%), 24 h after SKF96365 treatment were arrested in the G2/M phase. CCK-8 test showed that Eca109 ESCC cells received both SKF96365 and radiation showed the worst ability of cell proliferation. The same result was obtained in vivo. Nude mice received combined radio-SKF96365 showed the smallest tumor size and volume.
Conclusions: TRPC6 plays an important role in development of esophageal cancer, and SKF96365 may increase the sensitivity of radiotherapy. TRPC6 may become a new radiotherapy target in esophageal cancer.