共刺激分子B7-H1在食管癌细胞株中的表达及生物学功能

B7- HI expression and its contribution to the regulation of biological function in human esophageal cancer cell line

目的观察负性共刺激分子B7-H1在食管癌细胞株Eca-109中下调表达后对细胞生物学行为的影响。方法选取B7-H1阳性的食管癌细胞株Eca-109，通过RNA干扰技术下调B7-H1分子的表达并使用流式细胞术和Westernblot对其进行鉴定，采用细胞增殖实验、划痕实验和Transwell侵袭实验探讨B7-H1对食管癌细胞增殖、迁移及侵袭生物学行为的影响。结果通过RNA干扰技术成功构建B7-H1下调表达的食管癌细胞株Eca109-B7-H1-小干扰RNA（siRNA）及对照组Eca109-NC。细胞计数试剂盒（CCK-8）检测细胞增殖实验结果显示在培养的6、7、8d，Eca109-B7-H1-siRNA细胞相对增殖水平（0．61±0．03、0．78±0．04、0．89±0．04）显著低于Eca109-NC组（0．80±0．01、1．00±0．04、1．07±0．09），差异有统计学意义（P〈0．01），Eca109-NC组细胞与Eca-109组细胞间差异无统计学意义（P〉0．05）；划痕实验结果表明，划痕后24h，RNA干扰组细胞相对迁移距离（126．00±22．63）μm显著短于对照组（339．00±22．11）μm，差异有统计学意义（P〈0．05），且对照组与Eca-109细胞组间差异无统计学意义（P〉0．05）；Transwell侵袭实验结果表明，细胞接种24h后，干扰组的相对侵袭细胞数量[（20．60±4．61）个]显著少于对照组[（60．40±12．50）个]，差异有统计学意义（P〈0．01）。结论B7-H1在食管癌细胞株Eca-109中的下调表达可显著抑制细胞增殖、迁移及侵袭能力，食管癌细胞异常表达B7-H1参与调节其肿瘤生物学行为。

Objective To study the impact of downregulating the negative costimulation molecule B7 - H1 on biological behaviors in esophageal cancer cell line Eca - 109. Methods Down - regulation of B7 - H1 was performed by using RNAi method in the human esophageal cancer cell line Eca - 109. The ex- pression level of B7 - H1 was confirmed by using flow cytometry and Western blotting. The cell counting kit-8 （CCK- 8） assay, scratch assay and Transwell assay were used to investigate the contribution of 137 - H1 to biological behaviors of esophageal cancer cells. Results We successfully constructed stably re- duced expression of B7 - H1 in human esophageal cancer cell line by using RNAi named Eca109 - B7 - H1 - small interfering RNA （siRNA） and the control group Eca109 - NC. The CCK - 8 cell proliferation experiment showed that on the day 6, 7 and 8, the proliferation rate in Eca109 - B7 - H1 - siRNA group （0. 61 ± 0. 03, 0. 78 ± 0. 04, and 0. 89 ± 0.04） was significantly lower than in the control group （0. 80 ± 0. 01, 1.00 ± 0. 04, and 1.07 ±0. 09 ; P 〈 0. 01 ）, and there was no singnificant difference betweeen Eca109 - NC group and Eca - 109 group （P 〉 0. 05）. The scratch assay results revealed that the cell migra- tion ability in Eca109 - B7 - H1 - siRNA group [ （ 126. 00 ±22. 63 ） μm was significantly reduced as com- pared with the control group [ （339. 00±22. 11 ） μm] at 24 h （P 〈0. 05）. There was no singnificant differ- ence betweeen Eca109 - NC group and Eca - 109 group （P 〉 0. 05）. Transwell invasion assay results demon-strated that at 24 h after the cells were seeded, the number of invasive cells in Eca109 - B7 - H1 - siRNA group [ （20. 60±4. 61 ） cells] was singnificantly less than in the control group [ （60. 40 ± 12. 50） cells ,P 〈 0. 01]. Conclusion Down - regulation of B7 - H1 in esophageal cancer cell line Eca - 109 could signifi- cantly inhibit the cell proliferation, migration and invasion, suggesting that the abnormal expression of B7 - H1 in esophageal carcinoma cells could be involved in regulating the tumor biological behaviors and con- tribute to the tumor progression.