长链非编码RNA00461与原癌基因c-Myc结合调控三阴性乳腺癌细胞凋亡  

Long non-coding RNA00461 regulates apoptosis in triple-negative breast cancer cells by combining with the proto-oncogene c-Myc

作　　者：冯修来 刘慧芳 孙刚[1] Feng Xiulai;Liu Huifang;Sun Gang(Department of Breast and Thyroid Surgery,the Affiliated Cancer Hospital of Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学附属肿瘤医院乳腺甲状腺外科,乌鲁木齐830011

出　　处：《中华内分泌外科杂志》2023年第2期150-155,共6页Chinese Journal of Endocrine Surgery

基　　金：国家自然科学基金(82060520);新疆维吾尔自治区自然科学基金资助项目(2022D01D20、2020XS14)。

摘　　要：目的探索长链非编码RNA 00461(LINC00461)对三阴性乳腺癌(tripe negative breast cancer,TNBC)细胞凋亡的影响。方法用RT-qPCR检测正常乳腺上皮细胞MCF-10A与TNBC细胞系MDA-MB-231和MDA-MB-468细胞中LINC00461的表达。Annexin V-FITC/PI双染细胞凋亡试剂盒检测细胞凋亡率。Western blot法检测凋亡相关蛋白表达水平。FISH检测LINC00461在细胞中定位。RNA pulldown和RIP检测LINC00461与c-Myc的交互作用。裸鼠皮下成瘤实验验证LINC00461对TNBC凋亡水平的影响。结果与MCF-10A相比,MDA-MB-231和MDA-MB-468细胞中的LINC00461表达显著升高。干扰LINC00461显著降低LINC00461表达及c-Myc蛋白表达水平。过表达LINC00461显著降低凋亡相关蛋白表达水平。过表达LINC00461显著降低细胞凋亡率50%~70%,差异有统计学意义(P<0.05)。FISH结果显示LINC00461主要定位于细胞质。RNA pulldown结果显示LINC00461与c-Myc存在交互作用。RIP结果显示c-Myc组的LINC00461富集程度超过400%,差异有统计学意义(P<0.05)。此外过表达c-Myc可降低敲减LINC00461对细胞凋亡率30%~50%的促进作用。在动物实验中,过表达LINC00461显著提高的肿瘤体积(50%~80%)和肿瘤重量(30%~50%)。结论LINC00461调控TNBC细胞的凋亡水平,主要定位于细胞质,其分子作用机制可能是LINC00461与c-Myc蛋白发生交互作用从而发挥调控TNBC细胞凋亡的重要作用。Objective To investigate the effect of long non-coding RNA00461(LINC00461)on apoptosis of triple negative breast cancer cells.Methods RT-qPCR was used to detect the expression of LINC00461 in normal breast epithelial cells MCF-10A,TNBC cell lines MDA-MB-231 and MDA-MB-468.The apoptosis rate was detected by Annexin V-FITC/PI double staining cell apoptosis kit.Western blot was used to detect apoptosis-associated protein.The localization of LINC00461 in cells was detected by FISH.RNA pulldown and RIP were used to detect the interaction between LINC00461 and c-Myc.The effect of LINC00461 on TNBC apoptosis was verified by subcutaneous tumorigenesis in nude mice.Results Compared with MCF-10A,LINC00461 expression in MDA-MB-231 and MDA-MB-468 cells was significantly increased.Interference with LINC00461 significantly reduced the expression of LINC00461 and c-Myc protein.Overexpression of LINC00461 significantly decreased the expression level of apoptosis-related protein.Overexpression of LINC00461 significantly reduced the apoptosis rate by 50%-70%,with a statistically significant difference.FISH results showed that LINC00461 was mainly localized in the cytoplasm.RNA pulldown results showed that LINC00461 interacted with c-Myc.RIP results showed that the concentration of LINC00461 in c-Myc group was more than 400%,and the difference was statistically significant.In addition,overexpression of c-Myc reduced the promotion effect of knockdown LINC00461 on the apoptosis rate of 30%-50%.In animal experiments,overexpression of LINC00461 significantly increased tumor volume(50%-80%)and tumor weight(30%-50%).Conclusion LINC00461,mainly located in the cytoplasm,regulates the apoptosis of TNBC cells,and its molecular mechanism may be that LINC00461 interacts with c-Myc protein to play an important role in regulating the apoptosis of TNBC cells.

关 键 词：三阴性乳腺癌 长链非编码RNA00461 凋亡 C-MYC 

分 类 号：R737.9[医药卫生—肿瘤]