| 施凤涟,卫英,刘俊玲.miRNA-95在宫颈癌放疗敏感性中的作用及其机制研究[J].中华放射医学与防护杂志,2018,38(11):807-814 |
| miRNA-95在宫颈癌放疗敏感性中的作用及其机制研究 |
| The role of miRNA-95 in radiosensitivity of cervical cancer cells |
| 投稿时间：2018-04-11 |
| DOI：10.3760/cma.j.issn.0254-5098.2018.11.002 |
| 中文关键词: miRNA-95 宫颈癌 辐射敏感性 细胞增殖 细胞凋亡 |
| 英文关键词:miRNA-95 Cervical cancer Radiotherapy sensitivity Cell proliferation Apoptosis |
| 基金项目:河南省医学科技攻关支持项目基金（2014000157） |
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| 中文摘要: |
| 目的 研究miRNA-95在不同放射敏感性的宫颈癌患者肿瘤组织和宫颈癌细胞株中的表达情况及其调控表达后对宫颈癌细胞放射敏感性的影响。方法 分别利用Real-time PCR方法检测放疗敏感患者组20例、放疗耐受患者组20例的宫颈癌肿瘤组织和辐射抵抗宫颈癌细胞株（HeLa、SiHa）、辐射敏感宫颈癌细胞株（Me180）中miRNA-95的表达情况。利用脂质体2000将miRNA-95 mimics和miRNA-95 inhibitions转染至辐射抵抗的HeLa、SiHa细胞中，分别为miRNA-95mimics组和miRNA-95 inhibition组，同时设置miRNA-NC组为对照。CCk-8方法检测在0、2、4、6、8、10 Gy剂量60Co γ射线照射下各组宫颈癌细胞的增殖情况；平板单克隆实验检测4 Gy照射后各组宫颈癌细胞单克隆形成能力；流式细胞术检测4 Gy照射后各组宫颈癌细胞凋亡变化；双荧光素酶活性实验检测miRNA-95在宫颈癌细胞中的靶向基因；裸鼠实验检测4 Gy照射后各组宫颈癌裸鼠成瘤的变化。结果 miRNA-95在放疗耐受患者组宫颈癌肿瘤组织中表达显著高于放疗敏感组（t=12.279，P<0.05）；miRNA-95在HeLa、SiHa细胞中表达量显著，与Me180细胞比较，差异有统计学意义（t=5.162、7.114，P<0.05）；与miRNA-NC组相比，miRNA-95 inhibition组HeLa、SiHa细胞株中miRNA-95表达水平显著下降（t=9.284、8.036，P<0.05），而细胞增殖率显著下降（t=8.273、11.354、13.489、15.396和6.197、9.185、10.994、12.442，P<0.05）；单克隆形成率显著下降（t=8.378、7.931，P<0.05）；细胞凋亡率显著上升（t=10.265、8.386，P<0.05）； miRNA-95 inhibition组裸鼠成瘤重量显著降低（t=8.881、10.037，P<0.05）。结论 miRNA-95在放疗敏感的宫颈癌肿瘤组织和辐照敏感的宫颈癌细胞中均呈低表达，而抑制miRNA-95表达水平能够显著提高宫颈癌细胞的辐射敏感性，并通过靶向SGPP1基因从而发挥作用。 |
| 英文摘要: |
| Objective To detect the expression of miRNA-95 in cervical cancer tissues and cell lines with different radiosensitivity and study the effect of its regulation on radiosensitivity of cervical cancer cells. Methods Real-time PCR was used to detect the expression of miRNA-95 in cervical cancer tissues of 20 patients with radiosensitivity, 20 patients with radiation tolerance, radioresistant cervical cancer cell lines (HeLa, SiHa), and radiosensitive cervical cancer cell lines (Me180). MiRNA-95 mimics (miRNA-95 mimics group)and miRNA-95 inhibition (miRNA-95 inhibition group)were transfected into radioresistant HeLa and SiHa cells by liposome 2000, miRNA-NC was set as control group. CCk-8 assay was used to detect the proliferation of cervical cancer cells irradiated with 60Co γ-rays at 0,2,4,6,8,10 Gy. After 4 Gy irradiation, cell clonal formation ability was detected by plate monoclonal assay, and cell apoptosis was detected by flow cytometry. Dual luciferase activity assay was used to detect the target gene of miRNA-95 in cervical cancer cells. Nude mice were used to detect the changes of tumor formation ability. Results The expression of miRNA-95 in cervical cancer tissues of patients with radiotherapy tolerance was significantly higher than that of patients with radiotherapy sensitivity (t=12.279, P<0.05). The expressions of miRNA-95 in HeLa and SiHa cells were significantly higher those that of Me180 cells (t=5.162, 7.114, P<0.05). When the cells were treated with miRNA-95 inhibition, the expression of miRNA-95 in HeLa and SiHa cell lines was significantly lower than that of microRNA-NC group (t=5.162, 7.114, P<0.05), the cell proliferation rate decreased significantly (t=8.273, 11.354, 13.489, 15.396 and 6.197, 9.185, 10.994, 12.442, P<0.05), the cell monoclonal formation rate decreased significantly (t=8.378, 7.931, P<0.05), and the apoptosis rate increased significantly (t=10.265, 8.386, P<0.05). The tumorigenic weight of nude mice in the miRNA95 inhibition group was significantly decreased (t=8.881, 10.037, P<0.05). Conclusions The miRNA-95 had low levels in both radiosensitive cervical cancer tissues and cells. Inhibiting the expression of microRNA-95 can significantly improve the radiosensitivity of cervical cancer cells by targeting SGPP1 gene. |
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