| 吴 彬,陶 娟,佟 萌.miR-150-5p在甲状腺癌中的表达及其生物学功能研究[J].肿瘤学杂志,2019,25(6):503-509. |
| miR-150-5p在甲状腺癌中的表达及其生物学功能研究 |
| Expression and Biological Function of miR-150-5p in Thyroid Cancer |
| 投稿时间：2019-03-25 |
| DOI：10.11735/j.issn.1671-170X.2019.06.B004 |
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| 中文关键词: 甲状腺肿瘤 微小RNA-150 PI3K/Akt信号通路 增殖 凋亡 |
| 英文关键词:thyroid cancer mcrioRNA-150（miR-150-5p） PI3K/Akt signaling pathway prolife-ration apoptosis |
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| 中文摘要: |
| 摘 要：［目的］ 探讨微小RNA-150-5p（miR-150-5p）在甲状腺癌组织中的表达水平及其通过PI3K/Akt信号通路对甲状腺癌细胞增殖和凋亡能力的影响。［方法］ qRT-PCR检测68例甲状腺癌患者病变组织及其相应癌旁组织中miR-150-5p的表达水平。将miR-150-5p抑制剂和阴性对照转染至人甲状腺癌TPC-1细胞株，以空脂质体作为对照，qRT-PCR检测转染结果，MTT法检测细胞增殖能力，流式细胞术检测细胞凋亡，生物信息学网站Targetscan 预测miRNA-150-5p与PI3K的靶向结合情况，荧光素酶试验验证miR-150-5p与PI3K的靶向关系，Western blotting检测Cleaved caspase-3、PI3K、Akt、p-Akt蛋白的表达。［结果］ 68例甲状腺癌患者病变组织及相应癌旁组织中miR-150-5p的相对表达水平分别为0.96±0.06和1.66±0.11，差异有统计学意义（P＜0.05）；空白对照组和阴性对照组中miR-150-5p的表达水平差异无统计学意义（P＞0.05），miR-150-5p抑制剂组中miR-150-5p的相对表达量为0.12±0.02，明显低于空白对照组和阴性对照组（P＜0.05）。空白对照组和阴性对照组24、48和72h细胞增殖率的差异无统计学意义（P＞0.05），而miR-150-5p抑制剂组24、48和72h细胞增殖率分别为9.56%±1.38%、24.36%±4.66%和33.63%±5.32%，同一时间点均明显低于空白对照组和阴性对照组（P＜0.05）。空白对照组和阴性对照组48h细胞凋亡率的差异无统计学意义（P＞0.05），而miR-150-5p 抑制剂组的细胞凋亡率为16.25%±2.98%，明显高于空白对照组和阴性对照组（P＜0.01）。TargerScan网站预测结果显示，miR-150-5p可与PI3K互补结合，荧光素酶报告基因分析证实miR-150-5p与PI3K存在靶向结合位点。空白对照组和阴性对照组间PI3K、p-Akt和Cleaved caspase3蛋白表达的差异无统计学意义（P＞0.05），miR-150-5p抑制剂组PI3K、p-Akt蛋白表达低于空白对照组和阴性对照组（P＜0.01），Cleaved caspase3蛋白表达高于空白对照组和阴性对照组（P＜0.01），3组间Akt蛋白表达的差异无统计学意义（P＞0.05）。［结论］ miR-150-5p在甲状腺癌组织中表达异常升高，干扰miR-150-5p表达能够抑制甲状腺癌TPC-1细胞株的增殖，并通过上调Cleaved caspase-3促进细胞凋亡，其调控作用可能与PI3K/Akt信号通路有关。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the expression and biological function of microRNA-150-5p (miR-150-5p) in thyroid cancer. ［Methods］ The expression of miR-150-5p in cancer and adjacent tissues of 68 patients with thyroid cancer was detected by qRT-PCR. MiR-150-5p inhibitor and negative control were transfected into human thyroid cancer TPC-1 cells，and empty liposome was used as control. Transfection result was detected by qRT-PCR. After transfection，cell proliferation was determined by MTT，cell apoptosis as determined by flow cytometry. Targeted binding of miR-150-5p to PI3K was predicted through bioinformatics website TargetScan，and the targeting relationship between miR-150-5p and PI3K was verified by luciferase assay. Cleaved caspase-3，PI3K，Akt，p-Akt protein expression was determined by Western blotting.［Results］ The relative expression levels of miR-150-5p in the cancer and adjacent tissues were 0.96±0.06 and 1.66±0.11 respectively(P<0.05).The miR-150-5p expression in blank control group and negative control group was not significantly different(P>0.05)，and the relative expression of miR-150-5p in miR-150-5pinhibitor group was 0.12±0.02，significantly lower than that of blank control group and negative control groups(P<0.01). There were no significant differences in cell proliferation rates of miR-150-5p at 24，48 and 72h in blank and negative groups(P>0.05)，while those in miR-150-5p inhibitors group was 9.56%±1.38%，24.36%±4.66% and 33.63%±5.32% respectively，which were significantly lower than those in blank and negative control groups at the same time points(P<0.05). The apoptosis rates of blank and negative control groups had no significant different(P>0.05)，while the apoptosis rate of miR-150-5p inhibitors group was 16.25%±2.98%，significantly higher than that of blank control group and negative control group(P<0.01). TargerScan website predicted that microrna-150-5p might be complementary to PI3K，and luciferase reporter gene analysis confirmed targeted binding sites between microrna-150-5p and PI3K.There was no significant difference in the expression of PI3K，p-Akt and Cleaved caspase-3 protein between blank control group and negative control group(P>0.05). The protein expression of PI3K and p-AKT in miR-150-5p inhibitor group was significantly lower than that of blank and negative control groups(P<0.01)，and Cleaved caspase-3 protein was significantly higher than that of blank and negative control groups (P<0.01). The expression of Akt protein had no difference among the three groups(P>0.05). ［Conclusion］ MiR-150-5p is abnormally expressed in thyroid cancer tissues. The expression of miR-150-5p can significantly inhibit the proliferation of thyroid cancer TPC-1 cells，and promote cell apoptosis by raising cleaved caspase-3. Its regulative function may be associated with PI3K/AKT signal pathway. |
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