| 王海鹏,刘 屹,陈庆芬.三结构域家族蛋白22在宫颈癌中的表达及其对高危型人乳头瘤病毒阳性宫颈癌细胞增殖、侵袭及凋亡的影响[J].肿瘤学杂志,2021,27(9):758-765. |
| 三结构域家族蛋白22在宫颈癌中的表达及其对高危型人乳头瘤病毒阳性宫颈癌细胞增殖、侵袭及凋亡的影响 |
| Expression of Tripartite Motif Protein 22 in Cervical Cancer and Its Influence on Proliferation， Invasion and Apoptosis of Cervical Cancer Cells with High Risk Human Papillomavirus Infection |
| 投稿时间：2021-05-12 |
| DOI：10.11735/j.issn.1671-170X.2021.09.B011 |
|
 |
| 中文关键词: 宫颈肿瘤 乳头状瘤病毒 三结构域家族蛋白 细胞增殖 细胞凋亡 侵袭能力 |
| 英文关键词:cervical neoplasms papillomavirus tripartite motif protein cellular proliferation apoptosis invasion ability |
| 基金项目:国家自然科学基金（81402012）；陕西省自然科学研究基金（2015JQ8321，2019JM?鄄547）；西安交通大学基本业务项目（xyz012019112）；西安市科技计划项目［2019114613YX001SF035(3)］ |
|
| 摘要点击次数: 943 |
| 全文下载次数: 277 |
| 中文摘要: |
| 摘 要：［目的］ 探讨三结构域家族蛋白22（tripartite motif protein 22，TRIM22）在宫颈癌中的表达及其与临床病理学特征的关系，并分析TRIM22对高危型人乳头瘤病毒（human papilloma virus，HPV）阳性宫颈癌细胞增殖、侵袭和凋亡的影响。［方法］ 收集2017年8月至2018年9月41例手术切除或活检的宫颈癌组织和18例正常子宫颈组织，分别提取总RNA和总蛋白。实时定量PCR分析TRIM22 mRNA表达，Western blot分析TRIM22蛋白水平，并分析TRIM22 mRNA表达变化与患者临床病理学特征的关系。以HPV16阳性宫颈癌细胞系CaSki和HPV18阳性宫颈癌细胞系HeLa为研究对象，用TRIM22表达质粒（pUNO1-hTRIM22a）和对照表达质粒（pUNO1）稳定转染CaSki细胞和HeLa细胞，以未转染细胞为对照组。实时定量PCR和Western blot检测转染后细胞中TRIM22 mRNA表达和蛋白水平。采用细胞计数试剂盒-8（cell counting kit-8，CCK-8）检测细胞增殖；Transwell细胞侵袭实验检测细胞侵袭能力；流式细胞术检测细胞周期和凋亡；Western blot检测抗磷脂酰肌醇3激酶（phosphatidylinositol 3-kinase，PI3K）、蛋白激酶B（protein kinase B，AKT）和磷酸化AKT（phosphorylated AKT，p-AKT）蛋白表达。［结果］ TRIM22在宫颈癌组织中mRNA表达（0.99±0.16 vs 4.39±1.41，t=15.40，P<0.001）和蛋白水平（0.30±0.12 vs 1.32±0.27，t=19.97，P<0.001）均明显低于正常子宫颈组织，TRIM22在高危型HPV阳性宫颈癌组织中mRNA表达（0.94±0.14 vs 1.17±0.12，t=4.21，P<0.001）和蛋白水平（0.27±0.08 vs 0.43±0.18，t=3.86，P<0.001）均明显低于高危型HPV阴性宫颈癌组织。TRIM22 mRNA表达与淋巴结转移明显相关（1.04±0.16 vs 0.89±0.13，t=3.15，P=0.003），与肿瘤大小、临床分期无明显相关（P>0.05）。pUNO1转染CaSki细胞和HeLa细胞与未转染细胞在TRIM22 mRNA和蛋白水平、细胞增殖、侵袭数、细胞周期、凋亡率、PI3K、AKT、p-AKT蛋白表达量的差异均无统计学意义（P>0.05），pUNO1-hTRIM22a转染后CaSki细胞和HeLa细胞中TRIM22 mRNA和蛋白水平显著性高于pUNO1组和对照组（P<0.001），细胞增殖、侵袭数量（CaSki细胞：31.00±5.30 vs 135.30±17.24 vs 120.40±22.95，F=56.01，P<0.001；HeLa细胞：27.20±9.04 vs 112.80±12.03 vs 117.00±18.56，F=67.45，P<0.001）、处于G2~M期的细胞比例（CaSki细胞：2.04%±0.36% vs 12.72%±1.98% vs 12.40%±1.62%，F=57.82，P<0.001；HeLa细胞：3.07%±1.08% vs 8.26%±2.98% vs 10.92%±3.56%，F=22.82，P<0.001）、PI3K、p-AKT蛋白表达量均显著性低于pUNO1转染细胞和未转染细胞，而凋亡率显著性高于pUNO1组和对照组（CaSki细胞：11.90%±2.78% vs 5.27% ±1.78% vs 6.64%±1.19%，F=16.78，P<0.001；HeLa细胞：18.79%±4.64% vs 6.78%±1.03% vs 5.23%±1.09%，F=27.91，P<0.001）。［结论］ TRIM22在宫颈癌中的表达明显减低，其表达水平与高危型HPV感染、转移恶化密切相关。上调TRIM22表达可明显减弱高危型HPV阳性宫颈癌细胞的增殖、侵袭能力，促进宫颈癌细胞凋亡。 |
| 英文摘要: |
| Abstract： ［Objective］ To investigate the expression of tripartite motif protein 22(TRIM22) and its effects on proliferation， invasion and apoptosis of cervical cancer cells infected with high risk human papilloma virus(HPV). ［Methods］ Forty-one cervical cancer tissue samples and 18 normal cervical tissue samples were collected between August 2017 and September 2018. Total RNA and protein in tissue samples were extracted respectively. TRIM22 mRNA expression was analyzed by real-time RT-PCR， while TRIM22 protein level was analyzed by Western blot. The relationship between TRIM22 mRNA expression and clinical pathological features of patients was analyzed. The HPV16-positive cervical cancer CaSki cells and HPV18-positive cervical cancer HeLa cells were transfected with TRIM22 expression plasmid(pUNO1-hTRIM22a) and control plasmid(pUNO1)， the corresponding untransfected cells were used as control group. TRIM22 mRNA and protein expression was analyzed by qRT-PCR and Western blot， respectively. Cell proliferation was measured by cell counting kit-8(CCK-8)， cell invasion ability was detected by Transwell assay， cell cycle and apoptosis was determined by flow cytometry， the expression of phosphatidylinositol 3-kinase(PI3K)， protein kinase B(AKT) and phosphorylated AKT(p-AKT) was detected by Western blot. ［Results］ TRIM22 mRNA(0.99±0.16 vs 4.39±1.41， t=15.40， P<0.001) and protein level(0.30±0.12 vs 1.32±0.27， t=19.97， P<0.001) in cervical cancer tissues was significantly lower than those in normal cervical tissues. TRIM22 mRNA(0.94±0.14 vs 1.17±0.12， t=4.21， P=0.001) and protein level(0.27±0.08 vs 0.43±0.18， t=3.86， P<0.001) in high risk HPV-positive cervical cancer tissues was significantly lower than that in high risk HPV-negative cervical cancer tissues. TRIM22 mRNA expression was correlated with lymph node metastasis(1.04±0.16 vs 0.89±0.13， t=3.15， P=0.003)， however， did not correlate with tumor size or clinical stage(P>0.05). After transfection， TRIM22 mRNA and protein level was remarkably elevated in pUNO1-hTRIM22a group(P<0.001). Cell proliferation， cell migration(CaSki cells： 31.00±5.30 vs 135.30±17.24 vs 120.4±22.95， F=56.01， P<0.001， HeLa cells：27.20±9.04 vs 112.80±12.03 vs 117.0±18.56， F=67.45， P<0.001)， percentage of cells in G2~M stage(CaSki cells： 2.04%± 0.36% vs 12.72%±1.98% vs 12.40%±1.62%， F=57.82， P<0.001， HeLa cells： 3.07%±1.08% vs 8.26%±2.98% vs 10.92%±3.56%， F=22.82， P<0.001)， protein levels of PI3K and p-AKT in pUNO1-hTRIM22a group was significant lower， the percentage of apoptotic cells was significantly higher(CaSki cells： 11.90%±2.78% vs 5.27%± 1.78% vs 6.64%±1.19%， F=16.78， P<0.001， HeLa cells： 18.79%±4.64% vs 6.78%±1.03% vs 5.23%±1.09%， F=27.91， P<0.001) than those in pUNO1 group and control group. While there were no significant differences in TRIM22 mRNA or protein level， cell proliferation， invasion cell number， cell cycle， percentage of apoptotic cells， protein levels of PI3K， AKT or p-AKT between pUNO1 group and control group(P>0.05). ［Conclusion］ The expression of TRIM22 is significantly decreased in cervical cancer tissues. TRIM22 level is closely related to high risk HPV infection and metastasis. Up-regulation of TRIM22 expression can robustly attenuate proliferation and invasion ability and promote apoptosis of high risk HPV-positive cervical cancer cells. |
|
在线阅读
查看全文 查看/发表评论 下载PDF阅读器 |