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中华口腔医学研究杂志(电子版) ›› 2016, Vol. 10 ›› Issue (06) : 372 -376. doi: 10.3877/cma.j.issn.1674-1366.2016.06.002

所属专题: 文献

基础研究

低氧增强自噬促进舌鳞状细胞癌细胞迁移和侵袭
陈冠辉1, 王成1, 翁军权1, 梁建锋1, 李文清1, 竺越1, 侯劲松1,()   
  1. 1. 510055 广州,中山大学光华口腔医学院·附属口腔医院,广东省口腔医学重点实验室
  • 收稿日期:2016-10-12 出版日期:2016-12-01
  • 通信作者: 侯劲松
  • 基金资助:
    国家自然科学基金(81572660); 广东省自然科学基金(2015A030313034); 广东省科技计划(2013B021800059); 中山大学临床医学研究5010计划(2015018)

Hypoxia induced autophagy and promotes the migration and invasion in tongue squamous cell carcinoma

Guanhui Chen1, Cheng Wang1, Junquan Weng1, Jianfeng Liang1, Wenqing Li1, Yue Zhu1, Jinsong Hou1,()   

  1. 1. Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
  • Received:2016-10-12 Published:2016-12-01
  • Corresponding author: Jinsong Hou
  • About author:
    Corresponding author: Hiu Jinsong, Email:
引用本文:

陈冠辉, 王成, 翁军权, 梁建锋, 李文清, 竺越, 侯劲松. 低氧增强自噬促进舌鳞状细胞癌细胞迁移和侵袭[J]. 中华口腔医学研究杂志(电子版), 2016, 10(06): 372-376.

Guanhui Chen, Cheng Wang, Junquan Weng, Jianfeng Liang, Wenqing Li, Yue Zhu, Jinsong Hou. Hypoxia induced autophagy and promotes the migration and invasion in tongue squamous cell carcinoma[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2016, 10(06): 372-376.

目的

探讨低氧条件下对舌鳞状细胞癌(TSCC)自噬活性和迁移能力的影响。

方法

应用含1% O2的低氧培养箱培养TSCC UM1细胞,Western blot检测低氧诱导3、6、9、12、24 h后低氧诱导因子1α(HIF-1α)和自噬标记蛋白Beclin-1、自噬相关蛋白5(ATG5)和微管相关蛋白轻链3(LC3)的表达变化;细胞划痕实验检测低氧诱导后细胞迁移能力的改变;Transwell侵袭小室实验检测低氧诱导后细胞的侵袭能力;SPSS 13.0统计软件进行数据分析。

结果

低氧条件下,与对照组(0.527 ± 0.055)相比,TSCC UM1细胞自噬体双层膜标记蛋白LC3Ⅱ的表达(1.206 ± 0.053)增加(t= 12.96,P<0.001),同时自噬相关蛋白Beclin-1表达量(1.151 ± 0.078)较对照组(0.775 ± 0.062)明显提高(t= 6.736,P= 0.001),ATG5的表达(1.231 ± 0.06)较对照组(0.711 ± 0.052)也明显上升(t= 7.834,P= 0.001)。与对照组相比,低氧诱导后细胞迁移能力(0.349 ± 0.024)较对照组(0.788 ± 0.037)明显增加(t= 9.918,P= 0.001),细胞侵袭能力(23 ± 2)较对照组(71 ± 4)明显增强(t= 9.528,P= 0.001)。

结论

低氧条件下可以增强TSCC细胞自噬活性,促进其迁移和侵袭。

Objective

To investigate the effect of hypoxia on autophagy and migration of tongue squamous cell carcinoma (TSCC) cell line.

Methods

UM1 cells were cultured in hypoxia incubator containing 1% oxygen concentration. Western blot was performed to examine the expression of HIF-1α, Beclin-1, ATG5 and LC3 following 3, 6, 9, 12 and 24 h of treatment of hypoxia. Wounding healing was used to evaluate cell migration. Transwell assay was carried out to assess cell invasion. Statistical analysis was performed using SPSS 13.0 software.

Results

After, The expression level of autophagic protein LC3Ⅱ (1.206 ± 0.053) in the hypoxia treatment group was higher than control group (0.527 ± 0.055) and both showed statistically significant difference (t= 12.96, P<0.001) ; the expression level of Beclin-1 (1.151 ± 0.078) in the hypoxia treatment group was higher than control group (0.775 ± 0.062) , and both showed a statistically significant difference (t= 6.736, P= 0.001) ; the expression level of ATG5 (1.231 ± 0.06) in the hypoxia treatment group was higher than control group (0.711 ± 0.052) , was and both showed a statistically significant difference (t= 7.834, P= 0.001) . The migration ability of TSCCUM1 (0.349 ± 0.024) was improved under hypoxia in comparison to control groups (0.788 ± 0.037) (t= 9.918, P= 0.001) . The invasion ability of TSCCUM1 (23 ± 2) was upregulated under hypoxia in comparison to control groups (71 ± 4) (t= 9.528, P= 0.001) .

Conclusion

Hypoxia can improve autophagic activity levels and promotes the migration and invasion in tongue squamous cell carcinoma cell line.

图1 低氧处理不同时间后舌鳞状细胞癌细胞自噬相关蛋白表达变化
图2~5 低氧处理不同时间点后舌鳞状细胞癌细胞自噬相关蛋白相对表达变化
图6 低氧处理24 h后舌鳞状细胞癌UM1细胞迁移情况(× 50)
图7 低氧处理24 h后舌鳞状细胞癌UM1细胞划痕愈合率相对变化
图8 低氧处理后舌鳞状细胞癌细胞侵袭能力比较(× 100)
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