摘要
目的检测乳腺癌细胞放疗后LncRNA RP3-340B19.3的表达水平变化,探讨干预RP3-340B19.3后对乳腺癌细胞生物学特性的影响。方法采用实时荧光定量PCR(qRT-PCR)检测经辐照后乳腺癌细胞系MCF-7和MDA-MB-231中RP3-340B19.3的表达水平;利用小干扰RNA技术抑制两株细胞系中RP3-340B19.3的表达,qRT-PCR检测其转染效率;克隆形成试验检测RP3-340B19.3对乳腺癌细胞存活率的改变,并计算细胞放疗增敏比(SER);流式细胞术检测RP3-340B19.3对乳腺癌细胞的细胞凋亡与细胞周期的影响。结果qRT-PCR结果显示,经辐照后MCF-7和MDA-MB-231细胞中RP3-340B19.3的表达水平明显上调。转染siRNA后,MCF-7和MDA-MB-231细胞中RP3-340B19.3的相对表达量分别为0.2363±0.1538和0.3633±0.0371,均显著低于si-NC组(P<0.01)。经辐照后,RP3-340B19.3敲减组细胞克隆形成率和细胞存活分数明显下降(P<0.05),MCF-7和MDA-MB-231的细胞SER均显著升高。此外,经辐照后其细胞凋亡率显著上升(P<0.05);细胞周期发生改变,S期比例降低,细胞发生G_(0)/G_(1)期阻滞。结论乳腺癌细胞经辐照后RP3-340B19.3的表达水平明显升高,并且产生了放疗抗性。敲减RP3-340B19.3可以促进乳腺癌细胞的凋亡,抑制乳腺癌细胞的增殖,增强放射敏感性。
Objective To study the expression level of LncRNA RP3-340B19.3 in breast cancer cells after radiotherapy and the effects of interfering RP3-340B19.3 on the biological characteristics of breast cancer cells.Methods Quantitative Real-time PCR(qRTPCR)was used to detect the expression of RP3-340B19.3 in MCF-7 and MDA-MB-231 breast cancer cell lines after irradiation.The expression of RP3-340B19.3 was inhibited by small interfering RNA technique,and the transfection efficiency was detected by qRTPCR.Colony formation assay was used to detect the effect of RP3-340B19.3 on the survival rate of breast cancer cell lines,and the sensitizing enhancement ratio(SER)was calculated.The effects of RP3-340B19.3 on apoptosis and cell cycle of breast cancer cells were determined by flow cytometry.Results The expressions of RP3-340B19.3 in MCF-7 and MDA-MB-231 were up-regulated after irradiation.After transfection with si-RNA,the relative expression levels of RP3-340B19.3 in MCF-7 cells and MDA-MB-231 cells were 0.2363±0.1538 and 0.3633±0.0371,respectively,which were significantly lower than that in si-NC group(P<0.01).After irradiation,cell clonal formation rate and cell survival fraction in RP3-340B19.3 knockdown group were decreased significantly(P<0.05),the SER of MCF-7 and MDA-MB-231 was significantly elevated,respectively.The apoptosis rate of breast cancer cells was significantly increased(P<0.05).The cycle was changed,in which the proportion of S phase decreased and cell cycle arrest occurred in G_(0)/G_(1) phase.Conclusion The expression level of RP3-340B19.3 in breast cancer cells increased significantly after irradiation,which resulted in radiotherapy resistance.Knockdown of RP3 could promote the apoptosis of breast cancer cells,inhibit their proliferation and enhance radiosensitivity.
作者
吴茜茜
徐诗靓
赵玥昕
王凌霞
王波
杨欢
WU Xiai;XU Shiliang;ZHAO Yuexin;WANG Lingxia;WANG Bo;YANG Huan(Department of Clinical Laboratory,The Second Affiliated Hospital of Soochow University,Suzhou 215004,Jiangsu;Department of Oncology,The Second Affiliated Hospital of Soochow University,Suzhou 215004,Jiangsu;State Key Laboratory of Radiological Medicine and Radiation Protection,Soochow University,Suzhou 215004,Jiangsu,China)
出处
《临床检验杂志》
CAS
2023年第6期417-422,共6页
Chinese Journal of Clinical Laboratory Science
基金
国家自然科学基金青年项目(81702078)
江苏省自然科学基金(BK20170356)
苏州市科技计划项目(SKY2021045,SKJY2021097)
江苏省妇幼保健协会项目(FYX202123)
“姑苏卫生人才计划”青年拔尖人才项目
核技术医学应用重点人才项目(XKTJ-HRC2021001)
放射医学与辐射防护国家重点实验室资助项目(GZK1202137)
苏州市“科教兴卫”青年科技项目(KJXW2020019)
苏州市卫生青年骨干人才“全国导师制”项目(Qngg2023008)。
