肽基脯氨酰同分异构酶（Pin1）对子宫颈癌细胞脂质代谢的作用

郝立君; 徐丽秀; 李金秋; 马俊旗; 阿仙姑·哈斯木

doi:10.12259/j.issn.2095-610X.S20210111

肽基脯氨酰同分异构酶（Pin1）对子宫颈癌细胞脂质代谢的作用

doi: 10.12259/j.issn.2095-610X.S20210111

1.
新疆维吾尔自治区人民医院检验科，新疆乌鲁木齐　830001
2.
新疆医科大学基础医学院，新疆乌鲁木齐　830011
3.
新疆医科大学第一附属医院妇科，新疆乌鲁木齐　830011

基金项目: 国家自然科学基金资助项目（81960463）

详细信息

作者简介:
郝立君（1979～），男，新疆乌鲁木齐人，医学学士，副主任技师，主要从事临床医学检验工作

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: 2020-11-26
- 刊出日期: 2021-01-25

The Effect of Peptide-prolinyl Isomerase（Pin1）on Lipid Metabolism in Cervical Cancer Cells

1.
Dept. of Laboratory，People’s Hospital of Xinjiang Uygur Autonomous Region，Urumqi Xinjiang 830001
2.
Basic Medicine College of Xinjiang Medical University，Urumqi Xinjiang 830011
3.
Dept. of Gynecology，The 1st Affiliated Hospital of Xinjiang Medical University，Urumqi Xinjiang 830011，China

摘要

摘要: 目的探讨肽基脯氨酰同分异构酶（Pin1）蛋白对宫颈癌细胞脂质代谢的作用。方法应用免疫组织化学方法检测Pinl和ACC1蛋白在女性慢性宫颈癌及宫颈癌组织的表达；Western blotting技术和RT-PCR技术检测Pin1基因在宫颈癌SiHa、C33a及H8细胞中的蛋白本底表达情况，采用RNA干扰技术下调子宫颈癌C33a细胞中Pin1的表达；Western blotting技术检测宫颈癌细胞中Pin1蛋白表达水平变化对ACC1蛋白表达的影响；脂溶性荧光染色（BODIPY493/503）观察转染Pin1低表达慢病毒前后宫颈癌细胞中性脂肪含量的变化。结果与慢性宫颈炎相比，宫颈癌组织中Pin1和ACC1蛋白表达明显上调（P < 0.05），两者在宫颈癌组织中的表达呈正相关（ χ² = 6.02，P < 0.05）。与C33a细胞相比，SiHa细胞中Pin1蛋白的表达水平较低，故选用C33a细胞转染Pin1低表达慢病毒。C33a细胞转染Pin1低表达慢病毒后，Pin1及ACC1蛋白表达水平明显降低（ P < 0.05），与正常对照组和阴性对照组比，转染Pin1低表达慢病毒的细胞内中性脂肪酸含量降低。结论 Pin1蛋白的表达参与宫颈癌细胞内的脂质代谢，具体调控机制尚待研究。
- 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 peptide-based prolinyl isomerase（Pin1）protein on lipid metabolism in cervical cancer cells. Methods The expression of Pinl and ACC1 protein in chronic cervical cancer and cervical cancer tissues was detected by immunohistochemistry. Western blotting and RT-PCR were used to detect the background expression of Pin1 gene in SiHa, C33a and H8 cells of cervical cancer, and RNA interference was used to detect the expression of Pin1 in C33a cells of cervical cancer. The influence of Pin1 protein expression on ACC1 protein expression in cervical cancer cells was detected by Western blotting. Lipid-soluble fluorescence staining（bodipy493/503）was used to observe the changes in the content of neutral fat in cervical cancer cells before and after transfection with low expression of lentivirus Pin1. Results Compared with chronic cervicitis, the expression of Pin1 and ACC1 in cervical cancer tissues was significantly up-regulated（P < 0.05）, and the expression of Pin1 and ACC1 in cervical cancer tissues was positively correlated（r = 4.45, P < 0.05）. Compared with C33a cells, the expression level of Pin1 protein in SiHa cells was relatively low, so C33a cells were selected for transfection with lentivirus with low Pin1 expression. After the transfection with lentivirus with low Pin1 expression in C33a cells, the protein expression levels of Pin1 and ACC1 were significantly decreased（ P < 0.05）. Compared with normal control group and negative control group, the content of intracellular neutral fatty acids transfected with lentivirus with low Pin1 expression was decreased. Conclusion The expression of Pin1 protein is involved in lipid metabolism in cervical cancer cells, and the specific regulatory mechanism remains to be studied.
- Cervical cancer /
- Peptide-prolinyl isomerase /
- Fatty acid synthetase

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图 1 Pin1和ACC1蛋白在两种宫颈病变组织中的表达情况（×200）

A、B：宫颈癌组织；C、D：正常宫颈组织。

Figure 1. The expression of Pin1 and ACC1 protein in two cervical lesions

下载: 全尺寸图片幻灯片

图 2 Pin1蛋白在3株细胞中的表达情况

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 2. The expression of Pin1 protein in 3 cell lines

下载: 全尺寸图片幻灯片

图 3 Pin1 mRNA在3株细胞中的表达情况

A：Pin1的PCR结果；B：Pin1mRNA相对表达量。与正常宫颈上皮细胞比较，*P < 0.05。

Figure 3. The expression of Pin1 mRNA in 3 cell lines

下载: 全尺寸图片幻灯片

图 4 干扰Pin1后宫颈癌细胞中Pin1和ACC1蛋白表达情况

A：Pin、ACC的Western blotting结果；B：Pin、ACC蛋白相对表达量。与C33a细胞比较，^*P < 0.05。

Figure 4. The expression of Pin1 and ACC1 protein expression in cervical cancer cells after down regulate Pin1

下载: 全尺寸图片幻灯片

图 5 干扰Pin1基因后宫颈癌细胞内中性脂肪含量

A：shNON；B：shRACK1。

Figure 5. Neutral fat content in cervical cancer cells after down regulate Pin1

下载: 全尺寸图片幻灯片

表 1 Pinl和ACC蛋白在宫颈相应组织中的表达情况[n（%）]

Table 1. The expression of Pinl and ACC proteins in corresponding cervical tissues[n（%）]

分类	n	Pin1			ACC1
分类	n	阳性	χ²	P	阳性	χ²	P
慢性宫颈组织	30	9（30）	94.471	0.0000	11（36.67）	45.274	0.000
CSCC组织	64	59（92.18）^*			50（78.13）^*
与慢性宫颈炎组织比较，^*P < 0.05。

下载: 导出CSV

表 2 Pin1与ACC1表达水平的相关性分析

Table 2. Correlation analysis of expression levels of Pin1 and ACC1

Pin1	ACC1		χ²	P
Pin1	+	−	χ²	P
+	10	45	6.02	0.014
−	5	4

下载: 导出CSV

表 3 宫颈癌细胞中Pin1 mRNA的相对表达量（ $\bar x \pm s $ ）

Table 3. The relative expression of Pin1 mRNA in cervical cancer cells（ $\bar x \pm s $ ）

基因名	H8	SiHa	C33a	F	P
Pin1	0.17 ± 0.069*	1.0 ± 0.67	1.43 ± 0.26	91.997	0.000
与正常宫颈上皮细胞比较，*P < 0.05。

下载: 导出CSV

参考文献(12)

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