miR-128a介导的磷酸果糖激酶通过糖代谢促进非小细胞肺癌增殖的机制研究

doi:10.5246/jcps.2024.05.032

中国药学(英文版) ›› 2024, Vol. 33 ›› Issue (5): 430-437.DOI: 10.5246/jcps.2024.05.032

miR-128a介导的磷酸果糖激酶通过糖代谢促进非小细胞肺癌增殖的机制研究

张翔^*(), 应骏磊, 倪江伟

温州医科大学第一附属医院胸外科, 浙江温州 325000

收稿日期:2023-11-14 修回日期:2024-01-26 接受日期:2024-02-16 出版日期:2024-05-31 发布日期:2024-05-31
通讯作者: 张翔

Deciphering the mechanism of miR-128a-mediated proliferation of non-small cell lung cancer cells via phosphofructokinase and glycolysis

Xiang Zhang^*(), Junlei Ying, Jiangwei Ni

Thoracic Surgery Department, First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang, China

Received:2023-11-14 Revised:2024-01-26 Accepted:2024-02-16 Online:2024-05-31 Published:2024-05-31
Contact: Xiang Zhang
Supported by:
Wenzhou Science and Technology Bureau Science and Technology Plan Basic Research Project (Grant No. Y20190443).

摘要/Abstract

摘要：

探讨miR-128a介导的磷酸果糖激酶通过糖代谢促进非小细胞肺癌增殖的机制。采用生信分析方法预测与PFK作用的miRNA; 采用定量PCR方法 (qRT-PCR)方法在肿瘤组织和细胞样本上验证该分子表达量; 改变该分子表达量, 分别使用免疫荧光、Western blotting检测肺癌细胞的凋亡程度、PFK及低氧诱导因子1-α (hypoxia-inducible factor 1α, HIF1-α)表达水平。结果显示, 生信分析显示miR-128a可以靶向PFK; miR-128a在肺癌细胞中表达量明显降低, 过表达其水平, 细胞凋亡数目增多, 而PFR、HIF1-α蛋白表达水平降低。结果说明miR-128a在肺癌中表达降低, 通过降低PFK、HIF1-α表达促进了细胞增殖。

关键词: 非小细胞肺癌, miR-128a, 磷酸果糖激酶, 增殖

Abstract:

This study investigated the mechanism by which miR-128a, mediated through phosphofructokinase (PFK), fostered proliferation in non-small cell lung cancer (NSCLC) via glycolysis. Bioinformatics analysis was employed to predict microRNAs (miRNAs) involved in the interaction with PFK. Subsequently, the expression levels of the identified molecule were validated using qRT-PCR in both tumor tissues and cell samples. The modulation of this molecule’s expression was executed, and techniques such as immunofluorescence and Western blotting analysis were employed to evaluate apoptosis levels, PFK expression, and hypoxia-inducible factor-1 alpha (HIF1-α) expression in lung cancer cells. The results indicated that bioinformatics analysis revealed miR-128a as a potential targeting molecule for PFK. The expression level of miR-128a was significantly diminished in lung cancer cells. Overexpression of miR-128a led to increased apoptosis and reduced protein expression levels of PFK and HIF1-α. Conversely, decreased expression of miR-128a in lung cancer cells promoted cell proliferation by downregulating PFK and HIF1-α expression levels.

Key words: Non-small cell lung cancer, miR-128a, Phosphofructokinase, Proliferation

Supporting:

张翔, 应骏磊, 倪江伟. miR-128a介导的磷酸果糖激酶通过糖代谢促进非小细胞肺癌增殖的机制研究[J]. 中国药学(英文版), 2024, 33(5): 430-437.

Xiang Zhang, Junlei Ying, Jiangwei Ni. Deciphering the mechanism of miR-128a-mediated proliferation of non-small cell lung cancer cells via phosphofructokinase and glycolysis[J]. Journal of Chinese Pharmaceutical Sciences, 2024, 33(5): 430-437.

图/表 6

Table 1. Primer sequences.

Figure 1. Expression levels of miR-128a, PFK, and HIF1-α in NSCLC. (A) Detection of tumor tissue expression level; (B) Detection of tumor cell expression levels. *P < 0.05, **P < 0.01.

Figure 2. Verification of overexpression of miR-128a. (A) A549 cell line; (B) H460 cell line. *P < 0.05, **P < 0.01.

Figure 3. Effect of overexpression of miR-128a on live cell data: Green fluorescent cells represent survival (**P < 0.01).

Figure 4. The effect of overexpression of miR-128a on PFK expression (*P < 0.05).

Figure 5. The effect of overexpression of miR-128a on the expression of HIF1-α (*P < 0.05).

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miR-128a介导的磷酸果糖激酶通过糖代谢促进非小细胞肺癌增殖的机制研究

Deciphering the mechanism of miR-128a-mediated proliferation of non-small cell lung cancer cells via phosphofructokinase and glycolysis

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