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MiR-218-5p Inhibited the Stemness Maintaining and Invasion of A2B5+/CD133– Subgroup Human Glioma Stem Cells
Wu Zhiwu1#, Wang Hangzhou2#, Li Yanyan1, Li Xuetao1, Sun Ting1, Chen Guilin1, Xie Xueshun1, Zhou Youxin1*, Du Ziwei1
1Neurosurgery and Brain & Nerve Research Laboratory, the First Affiliated Hospital of Soochow University, Suzhou 215006, China; 2The Children’s Hospital of Soochow University, Suzhou 215003, China
Abstract: The article was aimed at detecting the expression of miR-218-5p in human gliomas， and exploring its influence in stemness maintaining and invasion of A2B5+/CD133– human glioma stem cells (SHG-139s). qRT-PCR was used to detect miR-218-5p expression in noncancerous brain tissue， human glioma tissue， human glioma cell lines and human glioma stem cell lines. Lentivirus vectors encoding miR-218-5p and antimiR-218-5p were constructed and stably transfected into A2B5+/CD133– SHG-139s cells. Neurosphere formation assay， Transwell assay， immunofluorescence and qRT-PCR analysis were used to explore the role of miR-218-5p in SHG-139s. qRT-PCR analysis showed that miR-218-5p expression was lower in human glioma tissues and cells than in noncancerous brain tissue and normal human astrocyte cells， and lower in A2B5+/CD133– (SHG-139s) cells than in CD133+ cells (SU2， U87s) cells. Immunofluorescence staining and qRT-PCR showed that miR-218-5p reduced the expressions of stem cell markers (A2B5， nestin， PLAGL2， ALDH1， Sox2)， compared to controls; However， qRT-PCR analysis showed that upregulated miR-218-5p expression led to a decline in CD133 expression， although this result was not confirmed by immunofluorescence staining. MiR-218-5p reduced SHG-139s cell invasiveness in Transwell assay and reduced MMP9 expression detected by qRT-PCR and immunofluorescence analysis. MiR-218-5p is implicated as a tumor suppressor gene in glioma that functions by upregulating miR-218-5p expression， which inhibits the stem cell and invasive properties of SHG-139s.
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