miR-148a-3p靶向调控PTEN基因表达对黑色素瘤细胞生物学特性的影响
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摘要
目的探讨miR-148a-3p靶向PTEN在黑色素瘤中的表达及其对黑色素瘤增殖、迁移及凋亡的影响。方法收集90例黑色素瘤癌组织及距肿瘤边缘5 cm的正常组织标本,记录患者临床病理资料,检测黑色素瘤组织及癌旁组织中miR-148a-3p、PTEN的表达水平;将miR-148a-3p、PTEN的siRNA转染人黑色素瘤SK-MEL-3细胞系,分别采用CCK8法、Transwell法和流式细胞术检测转染后细胞增殖水平、迁移能力、侵袭能力和凋亡情况。结果与癌旁正常组织相比,黑色素瘤组织中miR-148a-3p表达明显上调,PTEN表达明显下调(P<0.05)。miR-148a-3p表达水平与黑色素瘤淋巴结转移及TNM分期有关(P<0.05),PTEN表达与黑色素瘤淋巴结转移、肿瘤分化程度、TNM分期显著相关(P<0.05)。miR-148a-3p siRNA组PTEN基因的表达水平显著上调,细胞生长和迁移能力明显受到抑制,凋亡水平显著增加(P<0.05);miR-148a-3p siRNA+PTEN siRNA组细胞生长和迁移能力明显增强,细胞凋亡受到抑制(P<0.05)。结论 miR-148a-3p能负向调控PTEN的表达,从而促进黑色素瘤细胞的生长和迁移,并抑制其凋亡。
Objective To investigate miR-148 a-3 p in target regulating the expression of PTEN and its effects on cell proliferation,migration and apoptosis in melanoma. Methods The melanoma tissues and adjacent normal tissues 5 cm away from the tumor edge were collected from 90 cases of melanoma patients. Expression of miR-148 a-3 p and PTEN in both melanoma tissue and adjacent normal tissue were determined. Next, after siRNA of miR-148 a-3 p or PTEN was transfected into melanoma cell line SK-MEL-3, the cell proliferation,invasion, migration and apoptosis of SK-MEL-3 were detected using CCK-8 assay, Transwell assay and flow cytometry respectively. Results Initially, it was observed that miR-148 a-3 p was up-regulated while PTEN was down-regulated in melanoma tissues, when compared with adjacent normal tissues(P<0.05). Besides, expression of miR-148 a-3 p was related with melanoma lymph node metastasis and tumor node metastasis(TNM) stage(P<0.05). The PTEN expression was not only related with melanoma lymph node metastasis and TNM stage,but also tumor differentiation(P<0.05). In addition, miR-148 a-3 p siRNA elevated PTEN expression, and thus suppressed cell proliferation, invasion and migration and enhanced cell apoptosis in melanoma(P<0.05). After transfection of miR-148 a-3 p siRNA, PTEN siRNA promoted cell proliferation, invasion and migration and suppressed cell apoptosis in melanoma. Conclusion In conclusion, miR-148 a-3 p expression was negatively correlated with the PTEN expression. It may promote proliferation, invasion and migration, and inhibit apoptosis of SK-MEL-3 cells by down-regulating PTEN in melanoma.
Objective To investigate miR-148 a-3 p in target regulating the expression of PTEN and its effects on cell proliferation,migration and apoptosis in melanoma. Methods The melanoma tissues and adjacent normal tissues 5 cm away from the tumor edge were collected from 90 cases of melanoma patients. Expression of miR-148 a-3 p and PTEN in both melanoma tissue and adjacent normal tissue were determined. Next, after siRNA of miR-148 a-3 p or PTEN was transfected into melanoma cell line SK-MEL-3, the cell proliferation,invasion, migration and apoptosis of SK-MEL-3 were detected using CCK-8 assay, Transwell assay and flow cytometry respectively. Results Initially, it was observed that miR-148 a-3 p was up-regulated while PTEN was down-regulated in melanoma tissues, when compared with adjacent normal tissues(P<0.05). Besides, expression of miR-148 a-3 p was related with melanoma lymph node metastasis and tumor node metastasis(TNM) stage(P<0.05). The PTEN expression was not only related with melanoma lymph node metastasis and TNM stage,but also tumor differentiation(P<0.05). In addition, miR-148 a-3 p siRNA elevated PTEN expression, and thus suppressed cell proliferation, invasion and migration and enhanced cell apoptosis in melanoma(P<0.05). After transfection of miR-148 a-3 p siRNA, PTEN siRNA promoted cell proliferation, invasion and migration and suppressed cell apoptosis in melanoma. Conclusion In conclusion, miR-148 a-3 p expression was negatively correlated with the PTEN expression. It may promote proliferation, invasion and migration, and inhibit apoptosis of SK-MEL-3 cells by down-regulating PTEN in melanoma.
引文
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[8]Bhattacharya S, Chalk AM, Ng AJ, et al. Increased miR-155-5p and reduced miR-148a-3p contribute to the suppression of osteosarcoma cell death[J]. Oncogene, 2016,35(40):5282-5294. doi:10.1038/onc.2016.68.
[9]Bellissimo T, Russo E, Ganci F, et al. Circulating miR-21-5p and miR-148a-3p as emerging non-invasive biomarkers in thymic epithelial tumors[J]. Cancer Biol Ther, 2016,17(1):79-82. doi:10.1080/15384047.2015.1108493.
[10]Zhang H, Wang Y, Xu T, et al. Increased expression of micro RNA-148a in osteosarcoma promotes cancer cell growth by targeting PTEN[J]. Oncol Lett, 2016, 12(5):3208-3214.doi:10.3892/ol.2016.5050.
[11]Qingjuan L, Xiaojuan F, Wei Z, et al. miR-148a-3p overexpression contributes to glomerular cell proliferation by targeting PTEN in lupus nephritis[J]. Am J Physiol Cell Physiol, 2016, 310(6):C470-478. doi:10.1152/ajpcell.00129.2015.
[12]Edge SB, Compton CC. The American Joint Committee on Cancer:the 7th edition of the AJCC cancer staging manual and the future of TNM[J]. Ann Surg Oncol, 2010, 17(6):1471-1474. doi:10.1245/s10434-010-0985-4.
[13]郭军.黑色素瘤治疗研究进展[J].科技导报, 2014,(26):15-21. doi:10.3981/j. issn.1000-7857.2014.26.001.
[14]田红,肖桂芝,田苗,等.黑色素瘤治疗药物的研究进展[J].现代药物与临床, 2015, 7:890-896. doi:10.7501/j.issn.1674-5515.2015.07.030.
[15]陈勇,徐兴祥. miRNA-148a与肿瘤[J].肿瘤防治研究,2013, 40(3):301-303. doi:10.3971/j. issn.1000-8578.2013.03.019.
[16]马玉英,付冬晓,马微春. PTEN和PDCD4在胃癌组织中表达及与预后的关系[J].宁夏医学杂志, 2015, 1:19-21.doi:10.13621/j.1001-5949.2015.01.0019
[17]刘力山,刘丽霞,杨秋丽,等. PTEN和PDCD4在上皮性卵巢癌中的表达及意义[J].国际检验医学杂志, 2013,34(12):1527-1528. doi:10.3969/j. issn.1673-4130.2013.12.015.
[18]Liu CZ, Liu W, Zheng Y, et al. PTEN and PDCD4 are bona fide targets of microRNA-21 in human cholangiocarcinoma[J]. Chin Med Sci J, 2012, 27(2):65-72.
[19]宋润波,马建亭,孟增智,等.甲状腺乳头状癌中PTEN蛋白和肿瘤坏死因子的表达及意义[J].河北医药, 2015,37(18):2751-2753. doi:10.3969/j. issn.1002-7386.2015.18.009.
[2]杨跃.β-catenin对恶性黑色素瘤细胞A375的增殖、侵袭、迁移及凋亡的影响[D].南昌:南昌大学医学院,2014.
[3]Lucero OM, Dawson DW, Moon RT, et al. A re-evaluation of the"oncogenic"nature of Wnt/beta-catenin signaling in melanoma and other cancers[J]. Curr Oncol Rep, 2010,12(5):314-318. doi:10.1007/s11912-010-0114-3.
[4]成怀福,孙朝文,张广钰.人类Latexin基因与恶性肿瘤相关性的研究进展[J].医学理论与实践, 2018, 31(5):650-652. doi:10.19381/j. issn.1001-7585.2018.05.011.
[5]周明玉,岳双磊,何文杰,等. microRNA-214在恶性肿瘤中的表达及相关性的研究进展[J].现代生物医学进展, 2017, 17(4):787-790. doi:10.13241/j. cnki.pmb.2017.04.049.
[6]Wu T, Qu L, He G, et al. Regulation of laryngeal squamous cell cancer progression by the lncRNA H19/miR-148a-3p/DNMT1 axis[J]. Oncotarget, 2016, 7(10):11553-11566.doi:10.18632/oncotarget.7270.
[7]Wang X, Liang Z, Xu X, et al. miR-148a-3p represses proliferation and EMT by establishing regulatory circuits between ERBB3/AKT2/c-myc and DNMT1 in bladder cancer[J]. Cell Death Dis, 2016, 7(12):e2503. doi:10.1038/cddis.2016.373.
[8]Bhattacharya S, Chalk AM, Ng AJ, et al. Increased miR-155-5p and reduced miR-148a-3p contribute to the suppression of osteosarcoma cell death[J]. Oncogene, 2016,35(40):5282-5294. doi:10.1038/onc.2016.68.
[9]Bellissimo T, Russo E, Ganci F, et al. Circulating miR-21-5p and miR-148a-3p as emerging non-invasive biomarkers in thymic epithelial tumors[J]. Cancer Biol Ther, 2016,17(1):79-82. doi:10.1080/15384047.2015.1108493.
[10]Zhang H, Wang Y, Xu T, et al. Increased expression of micro RNA-148a in osteosarcoma promotes cancer cell growth by targeting PTEN[J]. Oncol Lett, 2016, 12(5):3208-3214.doi:10.3892/ol.2016.5050.
[11]Qingjuan L, Xiaojuan F, Wei Z, et al. miR-148a-3p overexpression contributes to glomerular cell proliferation by targeting PTEN in lupus nephritis[J]. Am J Physiol Cell Physiol, 2016, 310(6):C470-478. doi:10.1152/ajpcell.00129.2015.
[12]Edge SB, Compton CC. The American Joint Committee on Cancer:the 7th edition of the AJCC cancer staging manual and the future of TNM[J]. Ann Surg Oncol, 2010, 17(6):1471-1474. doi:10.1245/s10434-010-0985-4.
[13]郭军.黑色素瘤治疗研究进展[J].科技导报, 2014,(26):15-21. doi:10.3981/j. issn.1000-7857.2014.26.001.
[14]田红,肖桂芝,田苗,等.黑色素瘤治疗药物的研究进展[J].现代药物与临床, 2015, 7:890-896. doi:10.7501/j.issn.1674-5515.2015.07.030.
[15]陈勇,徐兴祥. miRNA-148a与肿瘤[J].肿瘤防治研究,2013, 40(3):301-303. doi:10.3971/j. issn.1000-8578.2013.03.019.
[16]马玉英,付冬晓,马微春. PTEN和PDCD4在胃癌组织中表达及与预后的关系[J].宁夏医学杂志, 2015, 1:19-21.doi:10.13621/j.1001-5949.2015.01.0019
[17]刘力山,刘丽霞,杨秋丽,等. PTEN和PDCD4在上皮性卵巢癌中的表达及意义[J].国际检验医学杂志, 2013,34(12):1527-1528. doi:10.3969/j. issn.1673-4130.2013.12.015.
[18]Liu CZ, Liu W, Zheng Y, et al. PTEN and PDCD4 are bona fide targets of microRNA-21 in human cholangiocarcinoma[J]. Chin Med Sci J, 2012, 27(2):65-72.
[19]宋润波,马建亭,孟增智,等.甲状腺乳头状癌中PTEN蛋白和肿瘤坏死因子的表达及意义[J].河北医药, 2015,37(18):2751-2753. doi:10.3969/j. issn.1002-7386.2015.18.009.
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