Pin1通过调控脂质代谢关键酶影响宫颈癌细胞的增殖及凋亡

海燕; 美力班·吐尔逊; 阿仙姑·哈斯木

doi:10.12259/j.issn.2095-610X.S20230107

Pin1通过调控脂质代谢关键酶影响宫颈癌细胞的增殖及凋亡

doi: 10.12259/j.issn.2095-610X.S20230107

新疆医科大学基础医学院/新疆地方病分子生物学实验室，新疆乌鲁木齐　836001

基金项目: 国家自然科学基金资助项目（81960463）；新疆维吾尔自治区天山创新团队基金资助项目（2021D14002）

详细信息

作者简介:
海燕（1996～），女，新疆焉耆人，在读硕士研究生，主要从事妇科肿瘤基础研究工作

通讯作者:
阿仙姑·哈斯木，E-mail：axiangu75@126.com

中图分类号: R737.33
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出版历程
- 收稿日期: 2022-10-08
- 网络出版日期: 2022-12-24
- 刊出日期: 2023-01-18

Effects of Pin1 on the Proliferation and Apoptosis of Cervical Cancer SiHa Cells by Regulating Key Enzymes of Lipid Metabolism

Basic Medical College of Xinjiang Medical University/Xinjiang Laboratory of Endemic Molecular Biology，Urumqi Xinjiang 836001，China

摘要

摘要: 目的探讨下调Pin1表达对宫颈癌SiHa细胞的脂质合成及增殖、凋亡的影响。方法 Western Blot检测Pin1在Hela、SiHa和H8细胞的表达情况；慢病毒转染技术构建Pin1低表达稳转细胞株，根据不同处理分为对照组（shPin1-NON）、敲低组1（shPin1-1）和敲低组2（shPin1-2），GEPIA2分析脂质代谢关键酶（ACC1、FASN）在宫颈癌及非肿瘤组织中的表达；Western Blot和qRT-PCR检测ACC1、FASN 蛋白和mRNA的表达；Western Blot检测Caspase-8、BCL-2、C-myc 蛋白表达；CCK-8检测细胞增殖能力；克隆形成实验检测细胞集落形成能力；流式细胞术检测细胞凋亡情况。结果 Pin1在宫颈癌细胞中高表达，且在SiHa细胞中表达最高（P < 0.05）；Pin1低表达慢病毒有效下调了Pin1表达水平；宫颈癌中ACC1、FASN mRNA表达高于非肿瘤组织；shPin1-1组和shPin1-2组ACC1、FASN 蛋白和mRNA表达均低于shPin1-NON组（P < 0.05）；shPin1-1组和shPin1-2组相较于shPin1-NON组Caspase-8蛋白表达被上调，而BCL-2、C-myc的表达均被抑制；shPin1-1组和shPin1-2组细胞增殖能力低于shPin1-NON组（P < 0.05）；shPin1-1组和shPin1-2组细胞集落形成能力小于shPin1-NON组（P < 0.05）。shPin1-1组和shPin1-2组细胞总凋亡率相较shPin1-NON组显著增加（P < 0.05）。结论 Pin1低表达有效下调宫颈癌SiHa细胞脂质代谢关键酶，抑制细胞增殖，诱发细胞凋亡，为靶向治疗宫颈癌提供新思路。
- 宫颈癌 /
- 肽基脯氨酰同分异构酶 /
- 脂肪酸合成酶
Abstract: Objective To investigate the effect of down-regulation of Pin1 expression on lipid synthesis, proliferation and apoptosis of cervical cancer SiHa cells. Methods Western Blot was used to detect the expression of Pin1 in Hela, SiHa and H8 cells; Lentiviral transfection technology constructed Pin1 low-expression stable transduction cell line, which was divided into the control group （shPin1-NON）, knockdown group1 （shPin1-1） and knockdown group2 （shPin1-2） according to different treatments; GEPIA2 was used to analyze the expression of key enzymes of lipid metabolism （ACC1, FASN） in cervical cancer and non-tumor tissues and the expression of ACC1 and FASN in cervical cancer and non-tumor tissues; Western Blot and qRT-PCR were used to detect the ACC1, FASN protein and mRNA expression; Western Blot was used to detect the Caspase-8, BCL-2, C-myc protein expression; CCK-8 was used to detect the cell proliferation; Clone formation assay was used to detect the cell colony formation ability and flow cytometry was used to detect the cell apoptosis. Results Pin1 was highly expressed in cervical cancer cells, and the highest expression was in SiHa cells; Pin1 low expression lentivirus effectively down-regulated the Pin1 expression level; ACC1 and FASN mRNA expressions in cervical cancer were higher than those in non-tumor tissues; The protein and mRNA expressions of ACC1 and FASN in shPin1-1 group and shPin1-2 group were lower than those in the shPin1-NON group; the shPin1-1 and shPin1-2 groups were up-regulated compared with the shPin1-NON group, while the expression of BCL-2 and C-myc was up-regulated. The expression was inhibited; The proliferation ability of shPin1-1 group and shPin1-2 group was lower than that of shPin1-NON group; The colony formation ability of shPin1-1 group and shPin1-2 group was lower than that of shPin1-NON group. Compared with the shPin1-NON group, the total apoptotic rate of cells in the shPin1-1 and shPin1-2 groups was significantly increased. （P < 0.05）. Conclusion Low expression of Pin1 can effectively down-regulate fatty acid synthase in cervical cancer SiHa cells, inhibit cell proliferation, and induce cell apoptosis, which provides a new idea for targeted therapy of cervical cancer.
- Cervical cancer /
- Peptidylprolyl isomerase /
- Fatty acid synthase

HTML全文

图 1 Pin1蛋白在宫颈癌及正常宫颈上皮的表达情况

A：Pin1的Western blotting结果;B:Pin1蛋白相对表达量。与正常宫颈上皮细胞比较，^*P < 0.05。

Figure 1. The Expression of Pin1 protein in cervical cancer and normal cervical epithelium

下载: 全尺寸图片幻灯片

图 2 检测Pin1慢病毒转染效率

A：转染慢病毒后Pin1蛋白的表达情况；B：Pin1蛋白相对表达量；C：转染慢病毒后Pin1的PCR结果。与shPin1-NON组比较，^*P < 0.05。

Figure 2. Detection of Pin1 lentiviral transfection efficiency

下载: 全尺寸图片幻灯片

图 3 shPin1对宫颈癌SiHa细胞脂质代谢关键酶的影响

A：GEPIA2 分析 ACC1、FASN 在宫颈癌中的表达；B：下调Pin1后 ACC1、FASN mRNA的表达情况；C：下调Pin1后ACC1、FASN 蛋白的表达情况；D：ACC1、FASN 蛋白相对表达量。与shPin1-NON组比较，^*P < 0.05。

Figure 3. The effect of shPin1 on key enzymes in lipid metabolism in SiHa cells of cervical cancer

下载: 全尺寸图片幻灯片

图 4 下调Pin1后宫颈癌SiHa细胞增殖和凋亡情况

A：下调Pin1后C-myc、BCL2、Caspase8 蛋白表达情况下；B：Pin1蛋白相对表达量；C：下调Pin1后SiHa细胞集落形成情况；D：下调Pin1后SiHa细胞增殖情况；E：下调Pin1后SiHa细胞凋亡情况。与shPin1-NON组比较，^*P < 0.05。

Figure 4. Proliferation and apoptosis of SiHa cells in cervical cancer after downregulation of Pin1

下载: 全尺寸图片幻灯片

表 1 qRT-PCR检测基因的引物序列

Table 1. Primer sequences of qRT-PCR detection genes

基因	引物序列（5′-3′）
Pin1	正向	CTGCCTTCAGCAGAGGTCAGATC
	反向	CAGTGCGGAGGATGATGTGGATG
ACC1	正向	TACCTTCTTCTACTGGCGGCTGAG
	反向	GCCTTCACTGTTCCTTCCACTTCC
FASN	正向	GTACACAGACAAAGCCCATTTT
	反向	TTTGGTTTACATCTGCACTTGG
β-actin	正向	TAGTTGCGTTACACCCTTTCTTG
	反向	TCACCTTCACCGTTCCAGTTT

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参考文献(17)

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