Department of Radiotherapy and Chemotherapy, The Second People's Hospital of Three Gorges University, Yichang 443000, Hubei Province, PR China; Department of Radiotherapy and Chemotherapy, The Second People's Hospital of Yichang, Yichang 443000, Hubei Province, PR China.
Department of Radiotherapy and Chemotherapy, The Second People's Hospital of Three Gorges University, Yichang 443002, Hubei Province, PR China; Discipline Inspection Office, The Second People's Hospital of Yichang, Yichang 443000, Hubei, PR China.
Exp Mol Pathol. 2019 Oct;110:104278. doi: 10.1016/j.yexmp.2019.104278. Epub 2019 Jun 24.
This present study is performed to figure out the role of microRNA-136 (miR-136) in radiosensitivity of esophageal squamous cell carcinoma (ESCC) cells through the regulation of MUC1.
Seventy-four ESCC patients were divided into radiotherapy sensitive group and radiotherapy resistance group. Colony formation assay and flow cytometry were used to test the radiosensitivity of radiotherapy resistant strain and parent strain. The expression of miR-136 between radiotherapy resistant strain and parent strain was detected by RT-qPCR, and the expression of miR-136 in Eca109 and TE-1 cells as well as Eca109-R and TE-1-R cells was detected after different doses of X-ray irradiation. Eca109 and TE-1 cells as well as Eca109-R and TE-1-R cells with overexpression of miR-136 or co-overexpression of miR-136 and MUC1 were constructed. Cell proliferation, colony formation and apoptosis was detected by CCK-8 assay, colony formation assay, and flow cytometry, respectively.
The expression of miR-136 in ESCC tissues was lower and MUC1 mRNA and protein expression was higher than that in adjacent normal tissues. The expression of miR-136 was negatively correlated with the expression of MUC1 mRNA in ESCC. Low expression of miR-136 and high expression of MUC1 were associated with tumor size, lymph node metastasis and distant metastasis. The expression of miR-136 increased while the expression of MUC1 decreased in the radiotherapy sensitive group of ESCC patients relative to the radiotherapy resistant group. The colony formation ability of radiation resistant cell line was stronger than that of parent cell line, and the apoptosis rate showed an opposite trend. Up-regulation of miR-136 reduced the survival rate, suppressed colony formation ability and induced apoptosis of ESCC cells under irradiation, which was reversed by upregulated MUC1.
This study demonstrates that up-regulation of miR-136 induces apoptosis and radiosensitivity of ESCC cells by inhibiting the expression of MUC1.
通过调节 MUC1，研究 miR-136 在食管鳞状细胞癌（ESCC）细胞放射敏感性中的作用。
将 74 例 ESCC 患者分为放疗敏感组和放疗抵抗组。集落形成实验和流式细胞术用于检测放疗抵抗株和亲本株的放射敏感性。通过 RT-qPCR 检测放疗抵抗株和亲本株之间 miR-136 的表达，检测不同剂量 X 射线照射后 Eca109 和 TE-1 细胞以及 Eca109-R 和 TE-1-R 细胞中 miR-136 的表达。构建 miR-136 过表达或与 MUC1 共过表达的 Eca109 和 TE-1 细胞以及 Eca109-R 和 TE-1-R 细胞。通过 CCK-8 检测、集落形成实验和流式细胞术分别检测细胞增殖、集落形成和细胞凋亡。
ESCC 组织中 miR-136 的表达较低，MUC1mRNA 和蛋白表达较高，与相邻正常组织相比。ESCC 中 miR-136 的表达与 MUC1mRNA 的表达呈负相关。miR-136 低表达和 MUC1 高表达与肿瘤大小、淋巴结转移和远处转移有关。与放疗抵抗组相比，ESCC 患者放疗敏感组 miR-136 的表达增加，而 MUC1 的表达降低。与亲本细胞系相比，辐射抵抗细胞系的集落形成能力更强，而凋亡率则呈现相反的趋势。上调 miR-136 可降低 ESCC 细胞在照射下的存活率，抑制其集落形成能力并诱导其凋亡，而上调 MUC1 则可逆转这一趋势。
本研究表明，上调 miR-136 通过抑制 MUC1 的表达诱导 ESCC 细胞凋亡和放射敏感性。
