B细胞淋巴瘤6对HTR-8/SVneo滋养细胞增殖、凋亡、侵袭和迁移的影响

doi:10.12280/gjfckx.20240555

国际妇产科学杂志 ›› 2024, Vol. 51 ›› Issue (6): 659-663.doi: 10.12280/gjfckx.20240555

B细胞淋巴瘤6对HTR-8/SVneo滋养细胞增殖、凋亡、侵袭和迁移的影响

杨丽, 杨静, 叶尔登切切克, 韩锐, 腊晓琳()

830011 乌鲁木齐，新疆医科大学第一附属医院生殖医学中心（杨丽，叶尔登切切克，韩锐，腊晓琳）；新疆医科大学第五附属医院妇科（杨静）；生殖疾病与出生缺陷综合防治研究中心，新疆生殖免疫临床医学研究中心，省部共建中亚高发病成因与防治国家重点实验室（腊晓琳）

收稿日期:2024-06-14 出版日期:2024-12-15 发布日期:2024-12-16
通讯作者: 腊晓琳，E-mail：xiaolinla@hotmail.com
基金资助:
中央引导地方科技发展资金项目(ZYYD2024ZY07);新疆维吾尔自治区研究生创新项目(XJ2024G152);新疆维吾尔自治区自然科学基金(2022D01C467);新疆维吾尔自治区卫生健康青年医学科技人才专项科研项目(WJWY-202410)

The Effect of BCL6 on Proliferation, Apoptosis, Invasion, and Migration of HTR-8/SVneo Trophoblast Cells

YANG Li, YANG Jing, YE Erdengqieqieke, HAN Rui, LA Xiao-lin()

Center for Reproductive Medicine, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, China (YANG Li, YE Erdengqieqieke, HAN Rui, LA Xiao-lin), Department of Gynecology, The Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China (YANG Jing); Research Center for Integrated Prevention and Treatment of Reproductive Diseases and Birth Defects, Xinjiang Clinical Research Center for Reproductive Immunology, State Key Laboratory of Pathogenesis, Prevention, and Treatment of High Incidence Diseases in Central Asia, Urumqi 830054, China (LA Xiao-lin)

Received:2024-06-14 Published:2024-12-15 Online:2024-12-16
Contact: LA Xiao-lin, E-mail: xiaolinla@hotmail.com

摘要/Abstract

摘要：

目的：探讨B细胞淋巴瘤6（B cell lymphoma 6，BCL6）基因过表达对人绒毛膜滋养细胞HTR-8/SVneo增殖、凋亡、侵袭、迁移的影响。方法：利用慢病毒介导的过表达LV5-BCL6（OE-BCL6）技术构建BCL6过表达的HTR-8/SVneo细胞系，同时设阴性对照组（OE-NC），并采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，qRT-PCR）和蛋白质印迹（Western blotting）检测BCL6的mRNA和蛋白表达水平。随后进行CCK8法、流式细胞术、Transwell实验以及Transwell小室侵袭实验，对比2组HTR-8/SVneo细胞增殖、凋亡、侵袭和迁移的能力。结果：荧光显微镜观察结果显示，OE-BCL6组和OE-NC组HTR-8/SVneo细胞的慢病毒转染后细胞生长状态良好，感染效率都达到了70%以上，慢病毒转染成功。mRNA水平检测显示，OE-BCL6组中BCL6表达高于OE-NC组（P<0.001）。蛋白水平检测显示，OE-BCL6组中BCL6表达高于OE-NC组（P<0.001）。转染48 h和72 h后，OE-BCL6组细胞的增殖活性较OE-NC组降低，OE-BCL6组细胞的凋亡率较OE-NC组升高（均P<0.05）。OE-BCL6组细胞侵袭和迁移的细胞数均较OE-NC组降低（均P<0.05）。结论：BCL6基因可抑制HTR-8/SVneo细胞增殖、侵袭及迁移，促进细胞凋亡，提示BCL6可能是临床病理妊娠诊疗的一个潜在靶点，但还需进一步研究阐明其分子机制。

关键词: BCL6, 人绒毛膜滋养细胞HTR-8/SVneo, 细胞增殖, 细胞凋亡, 侵袭

Abstract:

Objective: To explore the effects of overexpression of B cell lymphoma 6 (BCL6) gene on the proliferation, apoptosis, invasion, and migration of human trophoblast cells HTR-8/SVneo. Methods: Using lentivirus mediated overexpression of LV5-BCL6 (OE-BCL6) technology to construct HTR-8/SVneo cell lines with BCL6 overexpression, while a negative control group (OE-NC) was set up. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the mRNA and protein expression levels of BCL6. Subsequently, CCK8 assay, flow cytometry, Transwell assay, and Transwell chamber invasion assay were performed to compare the proliferation, apoptosis, invasion, and migration abilities of two groups of HTR-8/SVneo cells. Results: Fluorescence microscopy observation showed that the slow virus transfection of HTR-8/SVneo cells in the OE-BCL6 group and OE-NC group resulted in good cell growth and infection efficiency of over 70%, indicating successful slow virus transfection. mRNA level detection showed that BCL6 expression in the OE-BCL6 group was higher than that in the OE-NC group, and the difference was statistically significant (P<0.001). Protein level detection showed that the expression of BCL6 in the OE-BCL6 group was higher than that in the OE-NC group, and the difference was statistically significant (P<0.001). After transfection for 48 and 72 hours, the proliferation activity of OE-BCL6 group cells decreased compared to OE-NC group, while the apoptosis rate of OE-BCL6 group cells increased compared to OE-NC group, and the differences were statistically significant (all P<0.05). The amount of cells invaded and migratory cells in the OE-BCL6 group was significantly lower than that in the OE-NC group (both P<0.05). Conclusions: The BCL6 gene can inhibit the proliferation, invasion, and migration of HTR-8/SVneo cells, and promote cell apoptosis. BCL6 may be a potential target for diagnosis and treatment of clinical pathological pregnancy, but further research is needed to elucidate its molecular mechanism.

Key words: B cell lymphoma 6, Human trophoblast cells HTR-8/SVneo, Cell proliferation, Apoptosis, Invasion

杨丽, 杨静, 叶尔登切切克, 韩锐, 腊晓琳. B细胞淋巴瘤6对HTR-8/SVneo滋养细胞增殖、凋亡、侵袭和迁移的影响[J]. 国际妇产科学杂志, 2024, 51(6): 659-663.

YANG Li, YANG Jing, YE Erdengqieqieke, HAN Rui, LA Xiao-lin. The Effect of BCL6 on Proliferation, Apoptosis, Invasion, and Migration of HTR-8/SVneo Trophoblast Cells[J]. Journal of International Obstetrics and Gynecology, 2024, 51(6): 659-663.

图/表 4

图1 慢病毒感染效率及过表达效率（×200）注：A OE-NC组明视野，B OE-NC组荧光视野，C OE-BCL6组明视野，D OE-BCL6组荧光视野。

图2 慢病毒感染后BCL6 mRNA的表达水平

图3 慢病毒感染后BCL6蛋白表达水平

表1 2组HTR-8细胞增殖率比较（%）

组别	n	0 h	24 h	48 h	72 h
OE-NC组	3	100.00±1.46	100.01±7.91	100.00±2.68	100.00±1.37
OE-BCL6组	3	99.65±0.71	100.39±1.29	74.25±2.91	77.05±3.99
t		0.378	-0.083	11.287	9.417
P		0.725	0.938	<0.001	<0.001

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B细胞淋巴瘤6对HTR-8/SVneo滋养细胞增殖、凋亡、侵袭和迁移的影响

The Effect of BCL6 on Proliferation, Apoptosis, Invasion, and Migration of HTR-8/SVneo Trophoblast Cells

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