Relationship between DDX46 gene and invasion and metastasis behaviors of TE-1 esophageal squamous cell carcinoma cells_Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Authors：

LIN Junping , MENG Yuqi ,  LI Bin , FENG Haiming , LI Zheng

Department of Thoracic Surgery, Lanzhou University Second Hospital, Lanzhou University Second Clinical Medical College, Lanzhou, 730030, P. R. China;

Corresponding?author：

LI Bin, Email: dr.leebin@outlook.com

Keywords：

Esophageal squamous cell carcinoma; DDX46 gene; invasion; metastasis; integrin

DOI：

10.7507/1007-4848.202204019

Video：

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Objective To investigate the relationship between DDX46 genes and invasion and migration of esophageal squamous cell carcinoma cells. Methods Human esophageal squamous cell carcinoma cells TE-1 were transfected by fluorescent marker shRNA lentivirus (shDDX46 group), and an empty vector was transfected as a control (shCtrl group). The expression rate of green fluorescent protein under the microscope was used to evaluate the cell transfection efficiency. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blotting (WB) detected the knockdown efficiency of the target gene at the mRNA and protein expression levels. Wound healing, invasion assay and migration assay detected the changes of invasion and metastasis ability. Classical pathway analysis was used to explore signaling pathway changes and the possible mechanism of DDX46 in the invasion and metastasis was explored by detecting fibronectin expression. Results DDX46 gene at mRNA and protein levels was significantly inhibited after lentiviral transfection. Wound healing showed that after 8 h the cell mobility of TE-1 cells decreased significantly (P=0.001). Invasion assay showed that after 24 h the average cell metastasis rate of TE-1 cells was lower in the shDDX46 group than that in the shCtrl group (P<0.001). The cell metastasis rate in the shDDX46 group corresponding to observation points in the transwell assay was lower than that in the shCtrl group (P<0.001) after 24 h culture. The results of the classical pathway analysis showed that the integrin signaling pathway activity was inhibited, further exploration of the mechanism of action found that the expression of fibronectin associated with cell adhesion was decreased. Conclusion DDX46 gene is related to the invasion and migration ability of esophageal squamous cell carcinoma cells. Knockdown of DDX46 genes may reduce cell adhesion by downregulating the integrin pathway signaling.

Citation： LIN Junping, MENG Yuqi, LI Bin, FENG Haiming, LI Zheng. Relationship between DDX46 gene and invasion and metastasis behaviors of TE-1 esophageal squamous cell carcinoma cells. Chinese Journal of Clinical Thoracic and Cardiovascular Surgery, 2023, 30(7): 1030-1037. doi: 10.7507/1007-4848.202204019 Copy

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2.	焦曦, 魯智豪. 2021年食管癌臨床治療進展. 腫瘤綜合治療電子雜志, 2022, 8(1): 61-71.
3.	于振濤, 弓磊, 楊月陽, 等. 食管癌外科綜合治療進展. 中華消化外科雜志, 2022, 21(1): 30-33.
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6.	Li M, Ma Y, Huang P, et al. Lentiviral DDX46 knockdown inhibits growth and induces apoptosis in human colorectal cancer cells. Gene, 2015, 560(2): 237-244.
7.	Chen L, Xu M, Zhong W, et al. Knockdown of DDX46 suppresses the proliferation and invasion of gastric cancer through inactivating Akt/GSK-3β/β-catenin pathway. Exp Cell Res, 2021, 399(1): 112448.
8.	Jiang F, Zhang D, Li G, et al. Knockdown of DDX46 inhibits the invasion and tumorigenesis in osteosarcoma cells. Oncol Res, 2017, 25(3): 417-425.
9.	Lin Q, Jin HJ, Zhang D, et al. DDX46 silencing inhibits cell proliferation by activating apoptosis and autophagy in cutaneous squamous cell carcinoma. Mol Med Rep, 2020, 22(5): 4236-4242.
10.	Ma J, Gao ZY, Liu XN. DDX46 accelerates the proliferation of glioblastoma by activating the MAPK-p38 signaling. J Buon, 2021, 26(5): 2084-2089.
11.	李斌, 朱多杰, 何雯婷, 等. 靶向沉默DDX46基因對食管鱗癌TE-1細胞增殖及凋亡的影響. 中國胸心血管外科臨床雜志, 2017, 24(6): 463-468.
12.	李斌, 宋鐵牛, 蔣鵬, 等. DDX46基因對裸鼠食管鱗癌移植瘤生長的影響. 中國胸心血管外科臨床雜志, 2019, 26(2): 169-174.
13.	藺軍平. 干擾DDX46基因對食管鱗癌細胞侵襲、遷移能力的影響及機制探討. 蘭州大學, 2020.
14.	李斌, 李政, 尹泚, 等. 敲減DDX46后食管鱗癌細胞差異表達基因篩選及相互作用的生物信息學分析. 中國胸心血管外科臨床雜志, 2020, 27(1): 61-67.
15.	Kr?mer A, Green J, Pollard J, et al. Causal analysis approaches in ingenuity pathway analysis. Bioinformatics, 2014, 30(4): 523-530.
16.	You X, Cui H, Yu N, et al. Knockdown of DDX46 inhibits trophoblast cell proliferation and migration through the PI3K/Akt/mTOR signaling pathway in preeclampsia. Open Life Sci, 2020, 15(1): 400-408.
17.	Li Q, Lan T, Xie J, et al. Integrin-mediated tumorigenesis and its therapeutic applications. Front Oncol, 2022, 12: 812480.
18.	Bachmann M, Kukkurainen S, Hyt?nen VP, et al. Cell grins. Physiol Rev, 2019, 99(4): 1655-1699.
19.	黃夢汶, 林昶東, 陳劍峰. 整合素活化及其介導的黏著斑成熟與腫瘤轉移. 生理學報, 2021, 73(2): 149-159.
20.	Mishra YG, Manavathi B. Focal adhesion dynamics in cellular function and disease. Cell Signal, 2021, 85: 110046.
21.	Gahmberg CG, Gr?nholm M, Madhavan S. Regulation of dynamic cell adhesion by integrin-integrin crosstalk. Cells, 2022, 11(10): 1685.
22.	Pomella S, Cassandri M, Braghini MR, et al. New insights on the nuclear functions and targeting of FAK in cancer. Int J Mol Sci, 2022, 23(4): 1998.
23.	Li H, Gao Y, Ren C. Focal adhesion kinase inhibitor BI 853520 inhibits cell proliferation, migration and EMT process through PI3K/AKT/mTOR signaling pathway in ovarian cancer. Discov Oncol, 2021, 12(1): 29.
24.	Chuang HH, Zhen YY, Tsai YC, et al. FAK in cancer: From mechanisms to therapeutic strategies. Int J Mol Sci, 2022, 23(3): 1726.
25.	Owczarek C, Ortiz-Zapater E, Kim J, et al. CAR co-operates with integrins to promote lung cancer cell adhesion and invasion. Front Oncol, 2022, 12: 829313.
26.	Shi WK, Shang QL, Zhao YF. SPC25 promotes hepatocellular carcinoma metastasis via activating the FAK/PI3K/AKT signaling pathway through ITGB4. Oncol Rep, 2022, 47(5): 91.
27.	Luo J, Jiang Y, Wu L, et al. Long non-coding RNA ABHD11-AS1 promotes colorectal cancer progression and invasion through targeting the integrin subunit alpha 5/focal adhesion kinase/phosphoinositide 3 kinase/Akt signaling pathway. Aging (Albany NY), 2021, 13(16): 20179-20191.
28.	李斌, 藺軍平, 馮海明, 等. DDX46調控食管鱗癌TE-11細胞惡性生物學行為的機制探討. 中國癌癥雜志, 2018, 28(9): 671-678.
29.	吳瑞暖, 翁欣, 張萌, 等. PI3K/AKT信號通路抑制劑對食管癌細胞TE1的侵襲及遷移影響的機制研究. 深圳中西醫結合雜志, 2021, 31(5): 14-17.

1. Zheng RS, Zhang SW, Zeng HM, et al. Cancer incidence and mortality in China. 2016, JNCC, 2022, 2(1): 1-9.
2. 焦曦, 魯智豪. 2021年食管癌臨床治療進展. 腫瘤綜合治療電子雜志, 2022, 8(1): 61-71.
3. 于振濤, 弓磊, 楊月陽, 等. 食管癌外科綜合治療進展. 中華消化外科雜志, 2022, 21(1): 30-33.
4. Allemani C, Matsuda T, Di Carlo V, et al. Global surveillance of trends in cancer survival 2000-14 (CONCORD-3): Analysis of individual records for 37 513 025 patients diagnosed with one of 18 cancers from 322 population-based registries in 71 countries. Lancet, 2018, 391(10125): 1023-1075.
5. Weis K, Hondele M. The role of DEAD-Box ATPases in gene expression and the regulation of RNA-protein condensates. Annu Rev Biochem, 2022, 91: 197-219.
6. Li M, Ma Y, Huang P, et al. Lentiviral DDX46 knockdown inhibits growth and induces apoptosis in human colorectal cancer cells. Gene, 2015, 560(2): 237-244.
7. Chen L, Xu M, Zhong W, et al. Knockdown of DDX46 suppresses the proliferation and invasion of gastric cancer through inactivating Akt/GSK-3β/β-catenin pathway. Exp Cell Res, 2021, 399(1): 112448.
8. Jiang F, Zhang D, Li G, et al. Knockdown of DDX46 inhibits the invasion and tumorigenesis in osteosarcoma cells. Oncol Res, 2017, 25(3): 417-425.
9. Lin Q, Jin HJ, Zhang D, et al. DDX46 silencing inhibits cell proliferation by activating apoptosis and autophagy in cutaneous squamous cell carcinoma. Mol Med Rep, 2020, 22(5): 4236-4242.
10. Ma J, Gao ZY, Liu XN. DDX46 accelerates the proliferation of glioblastoma by activating the MAPK-p38 signaling. J Buon, 2021, 26(5): 2084-2089.
11. 李斌, 朱多杰, 何雯婷, 等. 靶向沉默DDX46基因對食管鱗癌TE-1細胞增殖及凋亡的影響. 中國胸心血管外科臨床雜志, 2017, 24(6): 463-468.
12. 李斌, 宋鐵牛, 蔣鵬, 等. DDX46基因對裸鼠食管鱗癌移植瘤生長的影響. 中國胸心血管外科臨床雜志, 2019, 26(2): 169-174.
13. 藺軍平. 干擾DDX46基因對食管鱗癌細胞侵襲、遷移能力的影響及機制探討. 蘭州大學, 2020.
14. 李斌, 李政, 尹泚, 等. 敲減DDX46后食管鱗癌細胞差異表達基因篩選及相互作用的生物信息學分析. 中國胸心血管外科臨床雜志, 2020, 27(1): 61-67.
15. Kr?mer A, Green J, Pollard J, et al. Causal analysis approaches in ingenuity pathway analysis. Bioinformatics, 2014, 30(4): 523-530.
16. You X, Cui H, Yu N, et al. Knockdown of DDX46 inhibits trophoblast cell proliferation and migration through the PI3K/Akt/mTOR signaling pathway in preeclampsia. Open Life Sci, 2020, 15(1): 400-408.
17. Li Q, Lan T, Xie J, et al. Integrin-mediated tumorigenesis and its therapeutic applications. Front Oncol, 2022, 12: 812480.
18. Bachmann M, Kukkurainen S, Hyt?nen VP, et al. Cell grins. Physiol Rev, 2019, 99(4): 1655-1699.
19. 黃夢汶, 林昶東, 陳劍峰. 整合素活化及其介導的黏著斑成熟與腫瘤轉移. 生理學報, 2021, 73(2): 149-159.
20. Mishra YG, Manavathi B. Focal adhesion dynamics in cellular function and disease. Cell Signal, 2021, 85: 110046.
21. Gahmberg CG, Gr?nholm M, Madhavan S. Regulation of dynamic cell adhesion by integrin-integrin crosstalk. Cells, 2022, 11(10): 1685.
22. Pomella S, Cassandri M, Braghini MR, et al. New insights on the nuclear functions and targeting of FAK in cancer. Int J Mol Sci, 2022, 23(4): 1998.
23. Li H, Gao Y, Ren C. Focal adhesion kinase inhibitor BI 853520 inhibits cell proliferation, migration and EMT process through PI3K/AKT/mTOR signaling pathway in ovarian cancer. Discov Oncol, 2021, 12(1): 29.
24. Chuang HH, Zhen YY, Tsai YC, et al. FAK in cancer: From mechanisms to therapeutic strategies. Int J Mol Sci, 2022, 23(3): 1726.
25. Owczarek C, Ortiz-Zapater E, Kim J, et al. CAR co-operates with integrins to promote lung cancer cell adhesion and invasion. Front Oncol, 2022, 12: 829313.
26. Shi WK, Shang QL, Zhao YF. SPC25 promotes hepatocellular carcinoma metastasis via activating the FAK/PI3K/AKT signaling pathway through ITGB4. Oncol Rep, 2022, 47(5): 91.
27. Luo J, Jiang Y, Wu L, et al. Long non-coding RNA ABHD11-AS1 promotes colorectal cancer progression and invasion through targeting the integrin subunit alpha 5/focal adhesion kinase/phosphoinositide 3 kinase/Akt signaling pathway. Aging (Albany NY), 2021, 13(16): 20179-20191.
28. 李斌, 藺軍平, 馮海明, 等. DDX46調控食管鱗癌TE-11細胞惡性生物學行為的機制探討. 中國癌癥雜志, 2018, 28(9): 671-678.
29. 吳瑞暖, 翁欣, 張萌, 等. PI3K/AKT信號通路抑制劑對食管癌細胞TE1的侵襲及遷移影響的機制研究. 深圳中西醫結合雜志, 2021, 31(5): 14-17.

Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Relationship between DDX46 gene and invasion and metastasis behaviors of TE-1 esophageal squamous cell carcinoma cells

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