RASSF6基因在食管癌细胞生长、增殖、侵袭、迁移中的细胞机制研究

Study on the cellular mechanism of RASSF6 gene in the growth,proliferation,invasion,and migration of esophageal cancer cells

目的探究RASSF6基因对食管癌细胞生长、增殖、侵袭、迁移的影响。方法回顾性收集2019年7月至2022年7月在新疆医科大学附属肿瘤医院接受治疗的200例食管癌患者的癌组织及癌旁组织,测定组织中RASSF6相对蛋白表达水平及mRNA水平。提取食管癌组织及癌旁组织细胞进行培养,敲除食管癌细胞中的RASSF6基因,构建缺失RASSF6食管癌细胞,检测正常食管癌细胞、癌旁组织细胞及缺失RASSF6食管癌细胞的生长周期、增殖、侵袭及迁移能力。结果食管癌组织中的RASSF6相对蛋白表达水平及mRNA水平均高于癌旁组织,差异均有统计学意义(P<0.05)。与癌旁组织细胞比较,食管癌细胞的G0/G1期所占比例降低,S期及G2/M期所占比例升高,凋亡率显著降低,差异均有统计学意义(P<0.05);与食管癌细胞比较,敲除RASSF6食管癌细胞的G0/G1期所占比例升高,S期及G2/M期所占比例降低,凋亡率显著升高,差异均有统计学意义(P<0.05)。与癌旁组织细胞及敲除RASSF6食管癌细胞比较,食管癌细胞培养24、48、72 h后的吸光度值显著升高,差异均有统计学意义(P<0.05);与食管癌细胞比较,敲除RASSF6食管癌细胞培养24、48、72 h后的吸光度值显著降低,差异均有统计学意义(P<0.05)。与癌旁组织细胞及敲除RASSF6食管癌细胞比较,食管癌细胞侵袭及迁移细胞数量显著升高,差异均有统计学意义(P<0.05);与食管癌细胞比较,敲除RASSF6食管癌细胞的侵袭及迁移细胞数量显著降低,差异均有统计学意义(P<0.05)。癌旁组织细胞和敲除RASSF6食管癌细胞的生长、增殖、侵袭、迁移能力相比,差异均无统计学意义(P>0.05)。结论与癌旁组织相比较,食管癌组织中RASSF6蛋白相对表达水平及mRNA水平均显著上调。敲除RASSF6基因可以显著抑制食管癌细胞的生长、增殖、侵袭、迁移能力。

Objective To explore the effect of RASSF6 gene on the growth,proliferation,invasion,and migration of esophageal cancer cells.Methods The cancer tissues and adjacent tissues of 200 esophageal cancer patients treated at the Cancer Hospital Affiliated to Xinjiang Medical University from July 2019 to July 2022 were retrospectively collected,and the relative protein expression and mRNA levels of RASSF6 in the tissues were measured.Esophageal cancer and adjacent tissue cells were extracted and cultured.The RASSF6 gene in esophageal cancer cells was knocked out,and RASSF6 deficient esophageal cancer cells were constructed.The growth cycle,proliferation,invasion,and migration ability of normal esophageal cancer cells,adjacent tissue cells,and RASSF6 deficient esophageal cancer cells were detected.Results The levels of RASSF6 relative protein and mRNA in esophageal cancer tissue were higher than those in adjacent tissues,the differences were statistically significant(P<0.05).Compared with adjacent tissue cells,the proportion of G0/G1 phase in esophageal cancer cells decreased,while the proportion of S phase and G2/M phase increased,and the apoptosis rate significantly decreased,the differences were statistically significant(P<0.05);compared with esophageal cancer cells,the proportion of G0/G1 phase in RASSF6 knockout esophageal cancer cells increased,while the proportion of S phase and G2/M phase decreased,and the apoptosis rate significantly increased,the differences were statistically significant(P<0.05).Compared with the adjacent tissue cells,the absorbance values of esophageal cancer cells significantly increased after 24,48,and 72 hours of culture,the differences were statistically significant(P<0.05);compared with esophageal cancer cells,the absorbance values of RASSF6 esophageal cancer cells cultured 24,48,and 72 hours after knocking out RASSF6 were significantly reduced,and the differences were statistically significant(P<0.05).Compared with adjacent tissue cells,the number of invasive and migratory cells in esophageal cancer cells significantly increased,the difference was statistically significant(P<0.05);compared with esophageal cancer cells,the number of invasive and migratory cells in knocking out RASSF6 significantly decreased,the difference was statistically significant(P<0.05).There was no statistically significant difference in the growth,proliferation,invasion,and migration abilities between adjacent cancer tissue cells and RASSF6 knockout esophageal cancer cells(P>0.05).Conclusion Compared with the adjacent tissue,the relative RASSF6 protein expression level and mRNA level were significantly up-regulated in esophageal cancer tissues.Knocking out the RASSF6 gene can significantly inhibit the growth,proliferation,invasion,and migration ability of esophageal cancer cells.