Knockdown of estrogen receptor alpha inhibits the proliferation and migration of A549 cells and the formation of transplanted tumors in nude mice_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

 CHEN Peirui ¹ , LV Jing ¹ , ZENG Chunfang ² , HE Xiaoyan ³ ,  DIAO Mingqiang ¹

1. Department of Cardiothoracic Surgery, Deyang People’s Hospital, Deyang, Sichuan 618000, P. R. China;
2. Department of Respiratory Medicine, Deyang People’s Hospital, Deyang, Sichuan 618000, P. R. China;
3. Department of Pathology, Deyang People’s Hospital, Deyang, Sichuan 618000, P. R. China;

Corresponding?author：

DIAO Mingqiang, Email: 1704981491@qq.com

Keywords：

Estrogen receptor α; A549 cells; Cell growth; Invasion; Tumor formation

DOI：

10.7507/1671-6205.202006060

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To explore the effect of interfering RNA (shRNA) on biological activity of A549 cells and tumor growth in nude mice after knockdown of estrogen receptor α (ERα) gene. Methods The ERα gene in A549 cells was knocked down by shRNA. RT-PCR and Western blot were used to detect the gene expression and protein expression after knockdown; colony formation experiment was used to detect the proliferation of cells, and RT-PCR was used to detect the expression of Ki-67 and PCNA; flow cytometry was used to detect apoptosis rate; transwell assay was used to detect cell invasion ability; Western blot was used to detect the expression of epithelial cadherin (E-cad) and neuropathic cadherin (N-cad) protein. The control group and A549 cells transfected with ERα-shRNA1 were injected subcutaneously in nude mice to construct transplanted tumors. Immunohistochemistry was used to detect the expression of Ki-67 and N-cad in tumor tissues. Results Compared with the control group, after transfection of ERα-shRNA1 and ERα-shRNA2, the mRNA and protein expressions of ERα were reduced significantly (P<0.05), and shRNA1 with high interference efficiency was used for subsequent experiments. Compared with the control group, the A549 cells were transfected with ERα-shRNA1, the colony formation rate was down-regulated significantly (P<0.05), the apoptosis rate was increased significantly (P<0.05), the expression of Ki-67 and PCNA were down-regulated significantly (P<0.05), the number of invasive cells was reduced significantly, the expression of E-cad was increased, and the expression of N-cad was decreased (P<0.05). The results of tumor formation in nude mice showed that interfering with ERα expression can significantly inhibit tumor growth (P<0.05), and down-regulate the rate of Ki-67 and N-cad positive cells (P<0.05). Conclusion Knockdown of ERα inhibits the proliferation and migration ability of NSCLC cells and the occurrence and development of transplanted tumors in nude mice.

Citation： CHEN Peirui, LV Jing, ZENG Chunfang, HE Xiaoyan, DIAO Mingqiang. Knockdown of estrogen receptor alpha inhibits the proliferation and migration of A549 cells and the formation of transplanted tumors in nude mice. Chinese Journal of Respiratory and Critical Care Medicine, 2021, 20(9): 649-655. doi: 10.7507/1671-6205.202006060 Copy

1.	Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin, 2016, 66(1): 7‐30.
2.	Schwartz AG, Wenzlaff AS, Prysak GM, et al. Reproductive factors, hormone use, estrogen receptor expression and risk of non-small cell lung cancer in women. J Clin Oncol, 2007, 25(36): 5785‐5792.
3.	MarquezGarban DC, Chen HW, Goodglick L, et al. Targeting aromatase and estrogen signaling in human non‐small cell lung cancer. Ann N Y Acad Sci, 2009, 1155(1): 194‐205.
4.	Zhang J, Guan X, Liang N, et al. Estrogen-related receptor alpha triggers the proliferation and migration of human non-small cell lung cancer via interleukin-6. Cell Biochem Funct, 2018, 36(5): 255-262.
5.	Dahlman-Wright K, Cavailles V, Fuqua SA, et al. International union of pharmacology. LXIV. estrogen receptors. Pharmacol Rev, 2006, 58(4): 773-781.
6.	Jameera Begam A, Jubie S, Nanjan MJ. Estrogen receptor agonists/antagonists in breast cancer therapy: a critical review. Bioorg Chem, 2017, 71: 257-274.
7.	Li Q, Gao H, Yang H, et al. Estradiol promotes the progression of ER+ breast cancer through methylation-mediated RSK4 inactivation. Onco Targets Ther, 2019, 12: 5907-5916.
8.	Che Q, Xiao X, Jun X, et al. 17beta-estradiol promotes endometrial cancer proliferation and invasion through IL-6 pathway. Endocr Connect, 2019, 8(7): 961-968.
9.	Frasor J, Danes JM, Komm B, et al. Estradiol promotes the progression up- and down-regulated gene expression in human breast cancer cells: insights into gene networks and pathways underlying estrogenic control of proliferation and cell phenotype. Endocrinology, 2003, 144(10): 4562-4574.
10.	顏浩, 徐英. 非小細胞肺癌中兩種雌激素受體亞型與血管內皮生長因子的表達及相關性研究. 中國呼吸與危重監護雜志, 2010, 9(3): 287-290.
11.	Zhao G, Zhao S, Wang T, et al. Estrogen receptor β signaling regulates the progression of Chinese non-small cell lung cancer. J Steroid Biochem Mol Biol, 2011, 124(1-2): 47-57.
12.	簡紅, 朱雷, 邵晉辰, 等. ERα、ERβ 在非吸煙 NSCLC 患者中的表達與生存期和性別的相關性研究. 中國癌癥雜志, 2013, 23(11): 910-916.
13.	王偉, 郭雯, 董海北, 等. 靈芝孢子提取物對肺腺癌細胞順鉑化療敏感性及 TAZ 表達的影響. 解放軍醫藥雜志, 2020, 32(6): 19-23.
14.	楊麗莎, 李福祥. 沉默 CDH1 基因促進非小細胞肺癌 A549 細胞系轉移侵襲的機制. 中國呼吸與危重監護雜志, 2019, 18(5): 452-456.
15.	呂薇, 龍波, 魏秀麗. miR-424 對肝癌細胞增殖、細胞周期和侵襲活性的影響. 解放軍醫藥雜志, 2020, 32(4): 33-35.
16.	徐敬軒, 楊巖, 張勖, 等. 柴胡皂苷 D 通過下調 miR-517a 調節膠質瘤細胞體外和體內移植瘤生長. 醫學分子生物學雜志, 2020, 17(1): 33-39.
17.	Tian W, Teng F, Gao J, et al. Estrogen and insulin synergistically promote endometrial cancer progression via crosstalk between their receptor signaling pathways. Cancer Biol Med, 2019, 16(1): 55-70.
18.	Arun A, Ansari MI, Popli P, et al. New piperidine derivative DTPEP acts as dual-acting anti-breast cancer agent by targeting ERalpha and downregulating PI3K/Akt-PKCα leading to caspase-dependent apoptosis. Cell Prolif, 2018, 51(6): e12501.
19.	許承斌, 丁明霞, 常媛媛, 等. 雌激素與非小細胞肺癌關系的研究進展. 醫學綜述, 2016, 22(21): 4215-4218.
20.	王紫翔, 白莎, 汪志國, 等. shRNA 干擾 lncRNA TUG1 對視網膜母細胞瘤生長的影響. 醫學分子生物學雜志, 2020, 17(4): 285-292.
21.	Zhang J, Li G, Li Z, et al. Estrogen-independent effects of ERα36 in ER-negative breast cancer. Steroids, 2012, 77(6): 666-673.
22.	Zhang S, Qiu C, Wang L, et al. The elevated level of ERα36 is correlated with nodal metastasis and poor prognosis in lung adenocarcinoma. Steroids, 2014, 87: 39-45.
23.	Liu DD, Zhang CJ, Li XL, et al. MicroRNA-567 inhibits cell proliferation, migration and invasion by targeting FGF5 in osteosarcoma. EXCLI J, 2018, 17: 102-112.
24.	Tong JS, Zhang QH, Wang ZB, et al. ER-α36, a novel variant of ER-α, mediates estrogen-stimulated proliferation of endometrial carcinoma cells via the PKCδ/ERK pathway. PLoS One, 2010, 5(11): e15408.
25.	Reinbolt RE, Mangini N, Hill JL, et al. Endocrine therapy in breast cancer: the neoadjuvant, adjuvant, and metastatic approach. Semin Oncol Nurs, 2015, 31(2): 146-155.
26.	Liu YF, Ma H, Yao J. ERα, a key target for cancer therapy: a review. Onco Targets Ther, 2020, 13: 2183-2191.

1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin, 2016, 66(1): 7‐30.
2. Schwartz AG, Wenzlaff AS, Prysak GM, et al. Reproductive factors, hormone use, estrogen receptor expression and risk of non-small cell lung cancer in women. J Clin Oncol, 2007, 25(36): 5785‐5792.
3. MarquezGarban DC, Chen HW, Goodglick L, et al. Targeting aromatase and estrogen signaling in human non‐small cell lung cancer. Ann N Y Acad Sci, 2009, 1155(1): 194‐205.
4. Zhang J, Guan X, Liang N, et al. Estrogen-related receptor alpha triggers the proliferation and migration of human non-small cell lung cancer via interleukin-6. Cell Biochem Funct, 2018, 36(5): 255-262.
5. Dahlman-Wright K, Cavailles V, Fuqua SA, et al. International union of pharmacology. LXIV. estrogen receptors. Pharmacol Rev, 2006, 58(4): 773-781.
6. Jameera Begam A, Jubie S, Nanjan MJ. Estrogen receptor agonists/antagonists in breast cancer therapy: a critical review. Bioorg Chem, 2017, 71: 257-274.
7. Li Q, Gao H, Yang H, et al. Estradiol promotes the progression of ER+ breast cancer through methylation-mediated RSK4 inactivation. Onco Targets Ther, 2019, 12: 5907-5916.
8. Che Q, Xiao X, Jun X, et al. 17beta-estradiol promotes endometrial cancer proliferation and invasion through IL-6 pathway. Endocr Connect, 2019, 8(7): 961-968.
9. Frasor J, Danes JM, Komm B, et al. Estradiol promotes the progression up- and down-regulated gene expression in human breast cancer cells: insights into gene networks and pathways underlying estrogenic control of proliferation and cell phenotype. Endocrinology, 2003, 144(10): 4562-4574.
10. 顏浩, 徐英. 非小細胞肺癌中兩種雌激素受體亞型與血管內皮生長因子的表達及相關性研究. 中國呼吸與危重監護雜志, 2010, 9(3): 287-290.
11. Zhao G, Zhao S, Wang T, et al. Estrogen receptor β signaling regulates the progression of Chinese non-small cell lung cancer. J Steroid Biochem Mol Biol, 2011, 124(1-2): 47-57.
12. 簡紅, 朱雷, 邵晉辰, 等. ERα、ERβ 在非吸煙 NSCLC 患者中的表達與生存期和性別的相關性研究. 中國癌癥雜志, 2013, 23(11): 910-916.
13. 王偉, 郭雯, 董海北, 等. 靈芝孢子提取物對肺腺癌細胞順鉑化療敏感性及 TAZ 表達的影響. 解放軍醫藥雜志, 2020, 32(6): 19-23.
14. 楊麗莎, 李福祥. 沉默 CDH1 基因促進非小細胞肺癌 A549 細胞系轉移侵襲的機制. 中國呼吸與危重監護雜志, 2019, 18(5): 452-456.
15. 呂薇, 龍波, 魏秀麗. miR-424 對肝癌細胞增殖、細胞周期和侵襲活性的影響. 解放軍醫藥雜志, 2020, 32(4): 33-35.
16. 徐敬軒, 楊巖, 張勖, 等. 柴胡皂苷 D 通過下調 miR-517a 調節膠質瘤細胞體外和體內移植瘤生長. 醫學分子生物學雜志, 2020, 17(1): 33-39.
17. Tian W, Teng F, Gao J, et al. Estrogen and insulin synergistically promote endometrial cancer progression via crosstalk between their receptor signaling pathways. Cancer Biol Med, 2019, 16(1): 55-70.
18. Arun A, Ansari MI, Popli P, et al. New piperidine derivative DTPEP acts as dual-acting anti-breast cancer agent by targeting ERalpha and downregulating PI3K/Akt-PKCα leading to caspase-dependent apoptosis. Cell Prolif, 2018, 51(6): e12501.
19. 許承斌, 丁明霞, 常媛媛, 等. 雌激素與非小細胞肺癌關系的研究進展. 醫學綜述, 2016, 22(21): 4215-4218.
20. 王紫翔, 白莎, 汪志國, 等. shRNA 干擾 lncRNA TUG1 對視網膜母細胞瘤生長的影響. 醫學分子生物學雜志, 2020, 17(4): 285-292.
21. Zhang J, Li G, Li Z, et al. Estrogen-independent effects of ERα36 in ER-negative breast cancer. Steroids, 2012, 77(6): 666-673.
22. Zhang S, Qiu C, Wang L, et al. The elevated level of ERα36 is correlated with nodal metastasis and poor prognosis in lung adenocarcinoma. Steroids, 2014, 87: 39-45.
23. Liu DD, Zhang CJ, Li XL, et al. MicroRNA-567 inhibits cell proliferation, migration and invasion by targeting FGF5 in osteosarcoma. EXCLI J, 2018, 17: 102-112.
24. Tong JS, Zhang QH, Wang ZB, et al. ER-α36, a novel variant of ER-α, mediates estrogen-stimulated proliferation of endometrial carcinoma cells via the PKCδ/ERK pathway. PLoS One, 2010, 5(11): e15408.
25. Reinbolt RE, Mangini N, Hill JL, et al. Endocrine therapy in breast cancer: the neoadjuvant, adjuvant, and metastatic approach. Semin Oncol Nurs, 2015, 31(2): 146-155.
26. Liu YF, Ma H, Yao J. ERα, a key target for cancer therapy: a review. Onco Targets Ther, 2020, 13: 2183-2191.

Chinese Journal of Respiratory and Critical Care Medicine

Knockdown of estrogen receptor alpha inhibits the proliferation and migration of A549 cells and the formation of transplanted tumors in nude mice

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content