2021 年第 10 期 第 16 卷

丙泊酚调控长链非编码RNA同源框基因反义RNA2对肾癌细胞生物学行为的影响

Effect of propofol on the biological behavior of renal cancer cells by regulating long non-coding RNA homeobox cluster antisense RNA2 

作者：丁容张文奇颜聪林冠文付强林慧欧阳碧山

英文作者：Ding Rong Zhang Wenqi Yan Cong Lin Guanwen Fu Qiang Lin Hui Ouyang Bishan

单位：海南省人民医院麻醉科，海口570311

英文单位：Department of Anesthesiology Hainan General Hospital Haikou 570311 China

关键词：肾癌；丙泊酚；同源框基因反义RNA2；细胞增殖；凋亡；迁移；侵袭

英文关键词：Renalcancer;Propofol;HomeoboxclusterantisenseRNA2;Cellproliferation;Apoptosis;Migration;Invasion

• 摘要：

• 目的  探讨丙泊酚对肾癌细胞生物学行为的影响及其潜在机制。方法以正常培养的人肾癌786-O细胞为对照组,用含2、5、10 mg/L丙泊酚的RPMI 1640培养液孵育的786-O细胞为2、5、10 mg/L丙泊酚组。应用细胞计数试剂盒法检测4组细胞活力，应用平板克隆实验检测4组细胞克隆形成能力，应用流式细胞仪检测4组细胞凋亡情况，应用Transwell实验检测4组细胞迁移和侵袭情况，应用实时定量聚合酶链反应法检测4组细胞同源框基因反义RNA2（HOXA-AS2）的表达，应用蛋白质印迹法检测B细胞淋巴瘤2家族蛋白（Bcl-2）和Bcl相关X蛋白（Bax）的表达情况。786-O细胞分别转染si-HOXA-AS2、si-NC、pcDNA-HOXA-AS2（si-HOXA-AS2组、si-NC组、pcDNA-HOXA-AS2组），用含10 mg/L丙泊酚的RPMI 1640培养液孵育转染pcDNA-HOXA-AS2细胞（丙泊酚+pcDNA-HOXA-AS2组），分别采用上述方法检测786-O细胞的活力、克隆形成、凋亡、迁移和侵袭能力。结果 2、5、10 mg/L丙泊酚组786-O细胞的吸光度值、克隆形成数、Bcl-2蛋白、HOXA-AS2的表达水平均显著低于对照组，细胞迁移数、侵袭数均显著少于对照组，凋亡率、Bax蛋白表达均显著多于对照组（均P<0.05）。随着丙泊酚浓度的增加，其对786-O细胞的吸光度值、克隆形成数、Bcl-2蛋白、HOXA-AS2表达以及细胞迁移和侵袭的抑制作用逐渐增强，对细胞凋亡、Bax蛋白表达的促进作用逐渐增强（均P<0.05）。si-HOXA-AS2组786-O细胞的吸光度值、克隆形成数、迁移数、侵袭数、Bcl-2蛋白和HOXA-AS2表达量均显著低于si-NC组，凋亡率、Bax蛋白表达量均显著高于si-NC组（均P<0.05）。丙泊酚+pcDNA-HOXA-AS2组786-O细胞HOXA-AS2的表达、吸光度值、克隆形成数、Bcl-2蛋白表达显著高于10 mg/L丙泊酚组［（0.840±0.079）比（0.153±0.017）、（0.77±0.04）比（0.33±0.03）、（98.0±2.4）比（40.7±2.0）、（0.623±0.037）比（0.173±0.012）］，细胞迁移数、侵袭数显著多于10 mg/L丙泊酚组［（145.7±7.6）个比（77.0±2.8）个、（116.3±3.7）个比（47.7±2.5）个］，凋亡率、Bax蛋白表达显著低于10 mg/L丙泊酚组［（11.25±0.69）%比（24.79±0.99）%、（0.300±0.029）比（0.820±0.071）］（均P<0.05）。结论 丙泊酚通过下调lncRNA HOXA-AS2的表达抑制肾癌细胞增殖、迁移和侵袭，并诱导细胞凋亡。

• Objective To investigate the effect of propofol on the biological behavior of renal cancer cells and its potential mechanism. Methods Normal cultured human renal cancer 786-O cells were used as control group, and  786-O cells incubated in RPMI 1640 medium containing 2, 5 and 10 mg/L propofol were used as 2, 5 and 10 mg/L propofol groups. In the four groups, cell viability was detected by cell counting kit method, cell colony forming ability was detected by colony forming cell assay, apoptosis was detected by flow cytometry, migration and invasion were detected by Transwell assay, homeobox cluster antisense RNA2(HOXA-AS2) was detected by real-time quantitative polymerase chain reaction (RT-PCR), and expressions of B cell lymphoma 2(Bcl-2) and Bcl-associated X protein(Bax) were detected by western blot. The 786-O cells were transfected with si-HOXA-AS2, si-NC and pcDNA-HOXA-AS2 (si-HOXA-AS2 group, si-NC group and pcDNA-HOXA-AS2 group). PcDNA-HOXA-AS2 cells were incubated with RPMI 1640 containing 10 mg/L propofol (propofol+pcDNA-HOXA-AS2 group). The viability, colony forming, apoptosis, migration and invasion of 786-O cells were detected by the above methods. Results The absorbance value, clone forming number, Bcl-2 protein and HOXA-AS2 expression levels of 786-O cells in 2, 5 and 10 mg/L propofol groups were lower that those in control group, the number of cell migration and invasion were less than those in control group, and the apoptosis rate and Bax protein expression were higher than those in control group (all P<0.05). With the increase of propofol concentration, the inhibitory effect of propofol on the absorbance value, colony forming number, Bcl-2 protein, HOXA-AS2 expressions, and cell migration and invasion of 786-O cells gradually increased, and the promoting effect on apoptosis and Bax protein expression gradually increased (all P<0.05). The absorbance value, clone forming number, migration number, invasion number, Bcl-2 protein and HOXA-AS2 expressions of 786-O cells in si-HOXA-AS2 group were lower than those in si-NC group, while the apoptosis rate and Bax protein expression were higher than those in si-NC group (all P<0.05). The HOXA-AS2 expression, absorbance value, clone forming number and Bcl-2 protein expression in propofol+pcDNA-HOXA-AS2 group were higher that those in 10 mg/L propofol group ［（0.840±0.079） vs （0.153±0.017）, （0.77±0.04） vs （0.33±0.03）,（98.0±2.4） vs （40.7±2.0）, （0.623±0.037） vs （0.173±0.012）］, cell migration number and invasion number were more than those in 10 mg/L propofol group ［（145.7±7.6） vs （77.0±2.8）, （116.3±3.7） vs （47.7±2.5）］, and apoptosis rate and Bax protein expression were lower that those in 10 mg/L propofol group ［（11.25±0.69）% vs （24.79±0.99）%, （0.300±0.029） vs （0.820±0.071）］(all P<0.05). Conclusion Propofol can inhibit the proliferation, migration and invasion of renal cancer cells and induce apoptosis by down regulating the expression of lncRNA HOXA-AS2.