Study on The Expression and Role of HOTAIR in CD133-Positive Gastric Cancer Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 XIXiao-long ^1,2 , JIANGBo-jian ¹ ,  YUJi-wei ¹

1. The First Department of General Surgery, Affiliated No. People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201900, China;
2. Graduate Department, Bengbu Medical College, Bengbu 233030, Anhui Province, China;

Corresponding?author：

YUJi-wei, Email: jiweiyu919@hotmail.com

Keywords：

Gastric cancer; Homeobox transcription antisense intergenic ribose nucleic acid; CD133; Small interfering RNA; Cell migration; Cell invasion

DOI：

10.7507/1007-9424.20150172

Video：

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Objective To study the expression and role of homeobox transcription antisense intergenic ribose nucleic acid (HOTAIR) in CD133-positive gastric cancer cells, which was classified to long non-coding RNA (LncRNA). Methods Immune magnetic cell sorting (MACS) was performed to sort CD133-positive and CD133-negative cells of KATO-Ⅲgastric cancer cells, then reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the expressions of HOTAIR mRNA and CD133 mRNA. After the intervention of small interfering RNA (siRNA) for CD133-positive KATO-Ⅲcells, RT-PCR method was performed to detect the expression of HOTAIR mRNA to select siRNA who had the best silent effect. The selected-siHOTAIR was used to silent the expression of HOTAIR, then the expressions of CD133 mRNA, E-cadherin mRNA, and N-cadherin mRNA were detected by RT-PCR. At last, Transwell experiments were performed to detect the migration ability and invasion ability. Results ①?RT-PCR test results showed that, the expression levels of CD133 mRNA and HOTAIR mRNA in CD133-positive group were significantly higher than those of CD133-negative group and no separation group (P < 0.05).②?After interference of siHOTAIR, the expression levels of HOTAIR mRNA in siHOTAIR1 group, siHOTAIR2 group, and siHOTAIR3 group were all significantly lower than those of blank control group and negative control group (P < 0.05), and the expression levels of HOTAIR mRNA in siHOTAIR2 group was lower than those of siHOTAIR1 group and siHOTAIR3 group (P < 0.05). The results indicated that siHOTAIR2 had the best interference efficiency.③?The expression levels of CD133 mRNA and N-cadherin mRNA in siHOTAIR2 group were lower than those corresponding indicators of blank control group and negative control group (P < 0.05), but the expression level of E-cadherin mRNA was higher than those of blank control group and negative control group (P < 0.05). Transwell experiment results showed that, number of cells which through the cell membrane in siHOTAIR2 group was lower than those of blank control group and negative control group (P < 0.05). Conclusion The expression of HOTAIR mRNA in CD133-positive KATO-Ⅲgastric cancer cells was higher than that of CD133-negative cells, interfering the expression of HOTAIR mRNA can reduce the expression of CD133 mRNA in CD133-positive KATO-Ⅲgastric cancer cells, and can inhibit cell migration and invasion.

Citation： XIXiao-long, JIANGBo-jian, YUJi-wei. Study on The Expression and Role of HOTAIR in CD133-Positive Gastric Cancer Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2015, 22(6): 659-664. doi: 10.7507/1007-9424.20150172 Copy

1.	?Jemal A, Bray F, Center MM, et al. Global cancer statistics. CA Cancer J Clin, 2011, 61(2):69-90.
2.	Hashimoto K, Aoyagi K, Isobe T, et al. Expression of CD133 in the cytoplasm is associated with cancer progression and poor prognosis in gastric cancer. Gastric Cancer, 2014, 17(1):97-106.
3.	Takaishi S, Okumura T, Tu S, et al. Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells, 2009, 27(5):1006-1020.
4.	Lee HH, Seo KJ, An CH, et al. CD133 expression is correlated with chemoresistance and early recurrence of gastric cancer. J Surg Oncol, 2012, 106(8):999-1004.
5.	蔡成, 俞繼衛, 吳巨鋼, 等. CD133通過上皮間質轉化促進胃癌侵襲和轉移的研究.中華胃腸外科雜志, 2013, 16(7):662-667.
6.	Li X, Wu Z, Fu X, et al. Long noncoding RNAs:insights from biological features and functions to diseases. Med Res Rev, 2013, 33(3):517-553.
7.	Gupta RA, Shah N, Wang KC, et al. Long non-coding RNA HOTAIR reprograms chromatin state to promote cancer metastasis. Nature, 2010, 464(7291):1071-1076.
8.	Kogo R, Shimamura T, Mimori K, et al. Long noncoding RNA HOTAIR regulates polycomb-dependent chromatin modification and is associated with poor prognosis in colorectal cancers. Cancer Res, 2011, 71(20):6320-6326.
9.	Kim K, Jutooru I, Chadalapaka G, et al. HOTAIR is a negative prognostic factor and exhibits pro-oncogenic activity in pancreatic cancer. Oncogene, 2013, 32(13):1616-1625.
10.	陸瑞祺, 吳巨鋼, 周國才, 等.胃癌CD133陽性細胞的純化及其生物學特性研究.中國普外基礎與臨床雜志, 2011, 18(12):1265-1270.
11.	Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med, 1997, 3(7):730-737.
12.	Iacopino F, Angelucci C, Piacentini R, et al. Isolation of cancer stem cells from three human glioblastoma cell lines:characterization of two selected clones. PLoS One, 2014, 9(8):e105166.
13.	Campos B, Herold-Mende CC. Insight into the complex regulation of CD133 in glioma. Int J Cancer, 2011, 128(3):501-510.
14.	Ferrandina G, Petrillo M, Bonanno G. Targeting CD133 antigen in cancer. Expert Opin Ther Targets, 2009, 13(7):823-837.
15.	Damdinsuren B, Nagano H, Kondo M, et al. TGF-beta1-induced cell growth arrest and partial differentiation is related to the suppression of Id1 in human hepatoma cells. Oncol Rep, 2006, 15(2):401-408.
16.	Angelastro JM, Lamé MW. Overexpression of CD133 promotes drug resistance in C6 glioma cells. Mol Cancer Res, 2010, 8(8):1105-1115.
17.	Han KS, Li N, Raven P, et al. Targeting integrin-linked kinase suppresses invasion and metastasis through downregulation of epithelial-to-mesenchymal transition in renal cell carcinoma. Mol Cancer Ther, 2015, 14(4):1024-1034.
18.	Shen W, Pang H, Liu J, et al. Epithelial-mesenchymal transition contributes to docetaxel resistance in human non-small cell lung cancer. Oncol Res, 2014, 22(1):47-55.
19.	Xu ZY, Yu QM, Du YA, et al. Knockdown of long non-coding RNA HOTAIR suppresses tumor invasion and reverses epithelial-mesenchymal transition in gastric cancer. Int J Biol Sci, 2013, 9(6):587-597.
20.	Yang F, Bi J, Xue X, et al. Up-regulated long non-coding RNA H19 contributes to proliferation of gastric cancer cells. FEBS J, 2012, 279(17):3159-3165.
21.	Sun M, Xia R, Jin F, et al. Downregulated long noncoding RNA MEG3 is associated with poor prognosis and promotes cell proliferation in gastric cancer. Tumour Biol, 2014, 35(2):1065-1073.
22.	Pádua Alves C, Fonseca AS, Muys BR, et al. Brief report:the lincRNA Hotair is required for epithelial-to-mesenchymal transition and stemness maintenance of cancer cell lines. Stem Cells, 2013, 31(12):2827-2832.
23.	Wu ZH, Wang XL, Tang HM, et al. Long non-coding RNA HOTAIR is a powerful predictor of metastasis and poor prognosis and is associated with epithelial-mesenchymal transition in colon cancer. Oncol Rep, 2014, 32(1):395-402.
24.	Fan Y, Shen B, Tan M, et al. Long non-coding RNA UCA1 increaseschemoresistance of bladder cancer cells by regulating Wnt signaling. FEBS J, 2014, 281(7):1750-1758.
25.	Wang G, Li Z, Zhao Q, et al. LincRNA-p21 enhances the sensitivity of radiotherapy for human colorectal cancer by targeting the Wnt/β-catenin signaling pathway. Oncol Rep, 2014, 31(4):1839-1845.

1. ?Jemal A, Bray F, Center MM, et al. Global cancer statistics. CA Cancer J Clin, 2011, 61(2):69-90.
2. Hashimoto K, Aoyagi K, Isobe T, et al. Expression of CD133 in the cytoplasm is associated with cancer progression and poor prognosis in gastric cancer. Gastric Cancer, 2014, 17(1):97-106.
3. Takaishi S, Okumura T, Tu S, et al. Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells, 2009, 27(5):1006-1020.
4. Lee HH, Seo KJ, An CH, et al. CD133 expression is correlated with chemoresistance and early recurrence of gastric cancer. J Surg Oncol, 2012, 106(8):999-1004.
5. 蔡成, 俞繼衛, 吳巨鋼, 等. CD133通過上皮間質轉化促進胃癌侵襲和轉移的研究.中華胃腸外科雜志, 2013, 16(7):662-667.
6. Li X, Wu Z, Fu X, et al. Long noncoding RNAs:insights from biological features and functions to diseases. Med Res Rev, 2013, 33(3):517-553.
7. Gupta RA, Shah N, Wang KC, et al. Long non-coding RNA HOTAIR reprograms chromatin state to promote cancer metastasis. Nature, 2010, 464(7291):1071-1076.
8. Kogo R, Shimamura T, Mimori K, et al. Long noncoding RNA HOTAIR regulates polycomb-dependent chromatin modification and is associated with poor prognosis in colorectal cancers. Cancer Res, 2011, 71(20):6320-6326.
9. Kim K, Jutooru I, Chadalapaka G, et al. HOTAIR is a negative prognostic factor and exhibits pro-oncogenic activity in pancreatic cancer. Oncogene, 2013, 32(13):1616-1625.
10. 陸瑞祺, 吳巨鋼, 周國才, 等.胃癌CD133陽性細胞的純化及其生物學特性研究.中國普外基礎與臨床雜志, 2011, 18(12):1265-1270.
11. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med, 1997, 3(7):730-737.
12. Iacopino F, Angelucci C, Piacentini R, et al. Isolation of cancer stem cells from three human glioblastoma cell lines:characterization of two selected clones. PLoS One, 2014, 9(8):e105166.
13. Campos B, Herold-Mende CC. Insight into the complex regulation of CD133 in glioma. Int J Cancer, 2011, 128(3):501-510.
14. Ferrandina G, Petrillo M, Bonanno G. Targeting CD133 antigen in cancer. Expert Opin Ther Targets, 2009, 13(7):823-837.
15. Damdinsuren B, Nagano H, Kondo M, et al. TGF-beta1-induced cell growth arrest and partial differentiation is related to the suppression of Id1 in human hepatoma cells. Oncol Rep, 2006, 15(2):401-408.
16. Angelastro JM, Lamé MW. Overexpression of CD133 promotes drug resistance in C6 glioma cells. Mol Cancer Res, 2010, 8(8):1105-1115.
17. Han KS, Li N, Raven P, et al. Targeting integrin-linked kinase suppresses invasion and metastasis through downregulation of epithelial-to-mesenchymal transition in renal cell carcinoma. Mol Cancer Ther, 2015, 14(4):1024-1034.
18. Shen W, Pang H, Liu J, et al. Epithelial-mesenchymal transition contributes to docetaxel resistance in human non-small cell lung cancer. Oncol Res, 2014, 22(1):47-55.
19. Xu ZY, Yu QM, Du YA, et al. Knockdown of long non-coding RNA HOTAIR suppresses tumor invasion and reverses epithelial-mesenchymal transition in gastric cancer. Int J Biol Sci, 2013, 9(6):587-597.
20. Yang F, Bi J, Xue X, et al. Up-regulated long non-coding RNA H19 contributes to proliferation of gastric cancer cells. FEBS J, 2012, 279(17):3159-3165.
21. Sun M, Xia R, Jin F, et al. Downregulated long noncoding RNA MEG3 is associated with poor prognosis and promotes cell proliferation in gastric cancer. Tumour Biol, 2014, 35(2):1065-1073.
22. Pádua Alves C, Fonseca AS, Muys BR, et al. Brief report:the lincRNA Hotair is required for epithelial-to-mesenchymal transition and stemness maintenance of cancer cell lines. Stem Cells, 2013, 31(12):2827-2832.
23. Wu ZH, Wang XL, Tang HM, et al. Long non-coding RNA HOTAIR is a powerful predictor of metastasis and poor prognosis and is associated with epithelial-mesenchymal transition in colon cancer. Oncol Rep, 2014, 32(1):395-402.
24. Fan Y, Shen B, Tan M, et al. Long non-coding RNA UCA1 increaseschemoresistance of bladder cancer cells by regulating Wnt signaling. FEBS J, 2014, 281(7):1750-1758.
25. Wang G, Li Z, Zhao Q, et al. LincRNA-p21 enhances the sensitivity of radiotherapy for human colorectal cancer by targeting the Wnt/β-catenin signaling pathway. Oncol Rep, 2014, 31(4):1839-1845.

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Study on The Expression and Role of HOTAIR in CD133-Positive Gastric Cancer Cells

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