南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (8): 1599-1604.doi: 10.12122/j.issn.1673-4254.2024.08.19
• • 上一篇
温小慧1,2(), 黄诗雅1,2, 刘学红1,2, 李坤寅3, 关永格3
收稿日期:
2024-01-24
出版日期:
2024-08-20
发布日期:
2024-09-06
通讯作者:
关永格
E-mail:18816781135@163.com
作者简介:
温小慧,博士,E-mail: 18816781135@163.com
基金资助:
Xiaohui WEN1,2(), Shiya HUANG1,2, Xuehong LIU1,2, Kunyin LI3, Yongge GUAN3
Received:
2024-01-24
Online:
2024-08-20
Published:
2024-09-06
Contact:
Yongge GUAN
E-mail:18816781135@163.com
摘要:
目的 基于Notch1信号观察子宫腺肌病(AM)在位内膜及Ishikawa细胞系糖酵解相关因子和蛋白的表达,并探讨子宫腺肌病的发生机制。 方法 收集AM(AM组,n=8)及子宫肌瘤(CON组,n=8)在位子宫内膜组织,通过临床特征评估两组子宫内膜的可比性, qRT-PCR和Western blot检测其Notch1信号及糖酵解相关因子和蛋白的表达,通过葡萄糖和乳酸试验检测其葡萄糖和乳酸的含量;建立过表达Notch1的Ishikawa细胞慢病毒稳转株, CCK-8检测其细胞存活率, Transwell迁移和侵袭实验检测其迁移和侵袭能力,细胞外酸化速率检测其糖酵解储备。 结果 与CON组子宫内膜组织比较,AM组糖类抗原125(CA125)水平明显高于CON组(P<0.01),AM患者在位子宫内膜组织Notch1、HK2和PDHA的mRNA相对表达量增加(P<0.01或P<0.05),Notch1、GLUT1、HK2、PKM和PDHA蛋白相对表达量均显著升高(P<0.01或P<0.05),葡萄糖含量显著下降、乳酸水平显著升高(P<0.01或P<0.05)。与对照病毒空载体(ov-NC)比较,过表达Notch1的Ishikawa细胞慢病毒稳转株(ov-Notch1)的细胞存活率显著升高(P<0.05),细胞迁移和侵袭细胞数量显著增加(P<0.05),酵解容量和酵解储备显著升高(P<0.01或P<0.05)。 结论 Notch1信号可能通过调控细胞的增殖、迁移、侵袭和糖酵解过程,进而参与AM的发生发展。
温小慧, 黄诗雅, 刘学红, 李坤寅, 关永格. Notch1信号通过调控细胞增殖、迁移、侵袭和糖酵解参与子宫腺肌病的发生及发展[J]. 南方医科大学学报, 2024, 44(8): 1599-1604.
Xiaohui WEN, Shiya HUANG, Xuehong LIU, Kunyin LI, Yongge GUAN. Role of Notch 1 signaling and glycolysis in the pathogenic mechanism of adenomyosis[J]. Journal of Southern Medical University, 2024, 44(8): 1599-1604.
Primer | Sequence (5' to 3') |
---|---|
Notch1 | TGGACCAGATTGGGGAGTTC |
GCACACTCGTCTGTGTTGAC | |
GLUT1 | TGGCATCAACGCTGTCTTCT |
AGCCAATGGTGGCATACACA | |
HK2 | GTGAATCGGAGAGGTCCCAC |
GCTAACTTCGGCCACAGGAT | |
PKM | ATGTCGAAGCCCCATAGTGAA |
TGGGTGGTGAATCAATGTCCA | |
PDHA | ATGGAATGGGAACGTCTGTTG |
CCTCTCGGACGCACAGGATA | |
GAPDH | CAGGAGGCATTGCTGATGAT |
GAAGGCTGGGGCTCATTT |
表1 引物序列
Tab.1 Primer sequences for RT-qPCR
Primer | Sequence (5' to 3') |
---|---|
Notch1 | TGGACCAGATTGGGGAGTTC |
GCACACTCGTCTGTGTTGAC | |
GLUT1 | TGGCATCAACGCTGTCTTCT |
AGCCAATGGTGGCATACACA | |
HK2 | GTGAATCGGAGAGGTCCCAC |
GCTAACTTCGGCCACAGGAT | |
PKM | ATGTCGAAGCCCCATAGTGAA |
TGGGTGGTGAATCAATGTCCA | |
PDHA | ATGGAATGGGAACGTCTGTTG |
CCTCTCGGACGCACAGGATA | |
GAPDH | CAGGAGGCATTGCTGATGAT |
GAAGGCTGGGGCTCATTT |
Group | Age (year) | Uterine volume (cm3) | Number of pregnancies | Number of abortions | CA125 (U/mL) |
---|---|---|---|---|---|
CON | 46.63±2.78 | 280.38 (128.75-1009.46) | 3.00 (2.00-3.00) | 0.00 (0.00-1.75) | 14.44 (12.16-23.82) |
AM | 44.88±2.95 | 636.08 (396.14-1020.82) | 3.50 (2.25-5.00) | 1.50 (0.25-3.50) | 45.01 (28.51-73.14)** |
表2 子宫内膜来源患者的基线特征
Tab.2 Baseline characteristics of the patients with adenomyosis and uterine fibroids [n=8, Mean±SD or M (P25-P75)]
Group | Age (year) | Uterine volume (cm3) | Number of pregnancies | Number of abortions | CA125 (U/mL) |
---|---|---|---|---|---|
CON | 46.63±2.78 | 280.38 (128.75-1009.46) | 3.00 (2.00-3.00) | 0.00 (0.00-1.75) | 14.44 (12.16-23.82) |
AM | 44.88±2.95 | 636.08 (396.14-1020.82) | 3.50 (2.25-5.00) | 1.50 (0.25-3.50) | 45.01 (28.51-73.14)** |
Group | Notch1 | GLUT1 | HK2 | PKM | PDHA |
---|---|---|---|---|---|
CON | 1.31±0.78 | 0.71 (0.65-1.13) | 1.01 (0.50-5.26) | 1.01 (0.29-3.63) | 1.78±1.68 |
AM | 5.12±3.46* | 3.04 (1.21-3.85) | 3.31 (1.58-12.38)* | 3.10 (1.40-8.02) | 6.92±5.05* |
表3 在位子宫内膜组织Notch1信号及糖酵解相关因子mRNA的表达
Tab.3 Relative mRNA expressions of Notch1 signaling pathway and glycolysis-related factors in the endometrial tissue [n=8, Mean±SD or M (P25-P75)]
Group | Notch1 | GLUT1 | HK2 | PKM | PDHA |
---|---|---|---|---|---|
CON | 1.31±0.78 | 0.71 (0.65-1.13) | 1.01 (0.50-5.26) | 1.01 (0.29-3.63) | 1.78±1.68 |
AM | 5.12±3.46* | 3.04 (1.21-3.85) | 3.31 (1.58-12.38)* | 3.10 (1.40-8.02) | 6.92±5.05* |
图1 在位子宫内膜组织Notch1信号及糖酵解相关蛋白的表达
Fig.1 Expression of Notch1 signal and glycolysis-related proteins in the endometrial tissues detected by Western blotting.
Group | Notch1 | GLUT1 | HK2 | PKM | PDHA |
---|---|---|---|---|---|
CON | 0.21±0.11 | 0.15±0.09 | 0.85±0.07 | 0.16±0.05 | 0.21±003 |
AM | 0.60±0.28** | 0.36±0.22* | 0.24±0.13* | 0.31±0.14* | 0.40±0.14* |
表4 在位子宫内膜组织Notch1信号及糖酵解相关蛋白的表达
Tab.4 Protein expressions of Notch1 signaling pathway and glycolysis-related factors in the endometrial tissue (n=7, Mean±SD)
Group | Notch1 | GLUT1 | HK2 | PKM | PDHA |
---|---|---|---|---|---|
CON | 0.21±0.11 | 0.15±0.09 | 0.85±0.07 | 0.16±0.05 | 0.21±003 |
AM | 0.60±0.28** | 0.36±0.22* | 0.24±0.13* | 0.31±0.14* | 0.40±0.14* |
Group | Glucose | Lactate |
---|---|---|
CON | 88.8±24.82 | 50.16±4.09 |
AM | 50.51±23.50* | 74.34±8.00** |
表5 在位子宫内膜组织葡萄糖、乳酸含量
Tab.5 Glucose and lactic acid levels in the endometrial tissue (n=5, Mean±SD)
Group | Glucose | Lactate |
---|---|---|
CON | 88.8±24.82 | 50.16±4.09 |
AM | 50.51±23.50* | 74.34±8.00** |
图2 过表达Notch1信号对Ishikawa细胞系迁移、侵袭的影响
Fig.2 Effects of overexpression of Notch1 signaling on migration and invasion of Ishikawa cells (Original magnification: ×200).
Group | Migration cells | Invasion cells |
---|---|---|
ov-NC | 4.78±1.68 | 9.89±4.00 |
ov-Notch1 | 64.56±17.67* | 140.89±48.49* |
表6 过表达Notch1信号对Ishikawa细胞系迁移、侵袭细胞数的影响
Tab.6 Effects of overexpression of Notch1 signaling on migration and invasion of Ishikawa cells (n=3, Mean±SD)
Group | Migration cells | Invasion cells |
---|---|---|
ov-NC | 4.78±1.68 | 9.89±4.00 |
ov-Notch1 | 64.56±17.67* | 140.89±48.49* |
Group | Fermentation capacity | Fermentation reserve |
---|---|---|
ov-NC | 10.43±1.27 | 9.61±1.88 |
ov-Notch1 | 17.13±1.68** | 16.20±1.62* |
表7 过表达Notch1信号对Ishikawa细胞系糖酵解能力的影响
Tab.7 Effect of overexpression of Notch1 signaling on glycolysis capacity of Ishikawa cells (n=3, Mean±SD)
Group | Fermentation capacity | Fermentation reserve |
---|---|---|
ov-NC | 10.43±1.27 | 9.61±1.88 |
ov-Notch1 | 17.13±1.68** | 16.20±1.62* |
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