苏丽萍, 季敏, 路子扬, 刘丽, 王维娜, 苗娜, 张巍, 蒲红伟. AJUBA调控MST1 YAP1和TAZ因子促进食管鳞状细胞癌细胞体外增殖 迁移和侵袭[J]. 中国肿瘤临床, 2022, 49(7): 331-337. DOI: 10.12354/j.issn.1000-8179.2022.20211829
引用本文: 苏丽萍, 季敏, 路子扬, 刘丽, 王维娜, 苗娜, 张巍, 蒲红伟. AJUBA调控MST1 YAP1和TAZ因子促进食管鳞状细胞癌细胞体外增殖 迁移和侵袭[J]. 中国肿瘤临床, 2022, 49(7): 331-337. DOI: 10.12354/j.issn.1000-8179.2022.20211829
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Liping Su, Min Ji, Ziyang Lu, Li Liu, Weina Wang, Na Miao, Wei Zhang, Hongwei Pu. Regulation of MST1, YAP1 and TAZ factors by AJUBA in proliferation, invasion and migration of esophageal squamous cell carcinoma[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2022, 49(7): 331-337. DOI: 10.12354/j.issn.1000-8179.2022.20211829
Citation: Liping Su, Min Ji, Ziyang Lu, Li Liu, Weina Wang, Na Miao, Wei Zhang, Hongwei Pu. Regulation of MST1, YAP1 and TAZ factors by AJUBA in proliferation, invasion and migration of esophageal squamous cell carcinoma[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2022, 49(7): 331-337. DOI: 10.12354/j.issn.1000-8179.2022.20211829
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AJUBA调控MST1 YAP1和TAZ因子促进食管鳞状细胞癌细胞体外增殖 迁移和侵袭

Regulation of MST1, YAP1 and TAZ factors by AJUBA in proliferation, invasion and migration of esophageal squamous cell carcinoma

    摘要
  • 摘要:
      目的  探讨AJUBA在食管鳞状细胞癌（esophageal squamous cell carcinoma，ESCC）细胞中表达以及是否通过调控MST1、YAP1和TAZ因子影响细胞增殖、侵袭和迁移。
      方法  采用Western blot 法检测AJUBA 蛋白在食管癌细胞系KYSE30、KYSE150与KYSE450中的表达水平，构建shRNA 干扰载体AJUBA转染至KYSE150食管癌细胞系；通过体外克隆形成实验、流式细胞术、划痕实验和Transwell实验探究AJUBA对KYSE150细胞的增殖、周期、迁移和侵袭能力的影响。采用RT-PCR和Western blot检测MST1、YAP1和TAZ mRNA和蛋白的表达；核质分离实验检测AJUBA、YAP1和TAZ蛋白表达情况。
      结果  稳定干扰AJUBA基因后，平板克隆实验结果显示，与阴性对照组相比，shAJUBA组的细胞克隆形成数量明显减少，差异具有统计学意义（P<0.001）；流式细胞周期实验结果显示，shAJUBA组中的细胞阻滞于G0/G1期，G2/M期与S期细胞显著减少（P<0.05）；划痕实验和Transwell侵袭实验结果显示，与AJUBA空染组相比，shAJUBA组细胞迁移能力和侵袭能力均明显减弱（P<0.001）；RT-PCR与Western blot实验结果显示，shAJUBA组细胞中的MST1、YAP1、TAZmRNA与蛋白表达水平均显著降低，差异具有统计学意义（P<0.05）。核质分离实验结果提示AJUBA蛋白在正常食管鳞状上皮细胞株（SHEE）细胞核中表达，而在KYSE150细胞胞质与胞核中表达；YAP1和TAZ蛋白在SHEE细胞中不表达，在KYSE150细胞质与细胞核中表达，上述结果差异均具有统计学意义（均P<0.05）。
      结论  AJUBA促进食管癌细胞的增殖、侵袭与迁移，可能与激活MST1、YAP1、TAZ因子的表达而影响食管鳞状细胞癌进展有关。

     

    Abstract:
      Objective  To investigate the expression of AJUBA in esophageal squamous cell carcinoma (ESCC) and whether AJUBA affects the proliferation, invasion, and migration of ESCC by regulating MST1, YAP1 and TAZ factors.
      Methods   Western blot was used to detect the expression levels of the AJUBA protein in the esophageal cancer cell lines KYSE30, KYSE150 and KYSE450. A ShRNA interference vector for AJUBA was constructed and transfected into the KYSE150 esophageal cancer cell lines. The effects of AJUBA on the proliferation, cycle, migration and invasion of KYSE150 cells were investigated by in vitro cloning and formation assay, flow cytometry, scratch assay, and Transwell migration assay. The mRNA and protein expressions of MST1, YAP1 and TAZ were detected by RT-PCR and Western blot. The expression of the AJUBA, YAP1 and TAZ proteins were detected by nucleo-cytoplasmic separation assay.
      Results  The stable knockdown of AJUBA gene in KYSE150 cell line was established. The results of plate cloning experiment showed that the number of cell clones formed in shAJUBA group was significantly lower than that in the negative control group (P< 0.0001). Flow cytometry results showed that KYSE150 cells in shAJUBA group were mainly blocked in G0/G1 phase, and and the number of cells were significantly reduced in G2/M and S phases (P<0.05). The results of scratch test and Transwell invasion test showed that the migration ability and invasion ability of shAJUBA group were significantly decreased compared with control group (P< 0.001). RT-PCR and Western blot results showed that the mRNA and protein expression levels of MST1, YAP1 and TAZ in shAJUBA group were significantly decreased, the differences were statistically significant (P<0.05). The results of nucleo-cytoplasmic separation indicated that AJUBA was expressed in SHEE (Human esophagus epithelial cell) nucleus, and AJUBA protein was expressed in KYSE150 cells cytoplasm and nucleus. The proteins of YAP1and TAZ were not expressed in SHEE cells, but were expressed in cytoplasm and nucleus of KYSE150 cells. The above results showed statistically significant differences (P<0.05).
      Conclusions  The AJUBA protein can promote the proliferation, invasion, and migration of esophageal carcinoma cells. This may be related to the activation of MST1, YAP1, and TAZ which may consequently affect the progression of ESCC.

     

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