沉默mGluR5对卵巢癌SKOV3细胞生物学特性的影响及其机制研究

目的 探讨小干扰RNA（siRNA）靶向代谢型谷氨酸受体5（mGluR5）对卵巢癌SKOV3细胞增殖、凋亡和迁移能力的影响及可能的机制。方法 采用靶向mGluR5表达的特异siRNA 转染卵巢癌SKOV3细胞（转染组），同时设置转染无义序列的阴性对照组。转染48 h，采用实时荧光定量PCR（QPCR）检测siRNA对mGluR5的沉默效果；采用CCK-8法、EdU细胞荧光染色实验、Hoechst 33342细胞染色实验、划痕实验检测沉默mGluR5表达后SKOV3细胞的活性、增殖、凋亡和迁移情况；采用Western blotting检测PTEN／Akt信号通路相关蛋白的表达情况。结果 QPCR检测显示，转染组中mGluR5 mRNA的表达水平降至（18.3±2.3）％，显著低于阴性对照组（P＜0.05）。CCK-8法检测显示，转染72 h阴性对照组的吸光值为4.1±0.2，高于转染组的2.4±0.3（P＜0.05）。EdU免疫荧光染色实验显示，转染48 h阴性对照组的细胞增殖率为（22.4±2.3）％，高于转染组的（9.2±1.2）％（P＜0.05）。Hoechst 33342细胞染色实验显示，转染48 h阴性对照组的细胞凋亡率为（6.2±1.3）％，低于转染组的（28.2±2.2）％（P＜0.05）。划痕实验显示，阴性对照组的相对划痕愈合比例为（100.0±2.1）％，显著高于转染组的（48.6±4.3）％（P＜0.05）。Western blotting检测显示，转染组PTEN蛋白的表达增加，p-Akt蛋白的表达降低（P＜0.05）；而Akt蛋白无明显变化（P＞0.05）。结论 沉默mGluR5表达能显著抑制卵巢癌SKOV3细胞的增殖，促进凋亡，降低细胞迁移能力，该过程可能与激活PTEN／Akt信号通路有关。

Study of metabotropic Glutamate receptor 5 silence on biological behaviors of ovarian cancer SKOV3 cells and possible mechanism

Objective To explore the effect of small interfering RNA (siRNA) targeting metabotropic Glutamate receptor 5 (mGluR5)on the biological behaviors of ovarian cancer SKOV3 cells as well as possible mechanism.Methods siRNA designed for specific targeting mGluR5 were transfected into SKOV3 cells as transfection group.Meanwhile,antisense sequence was used as negative control (NC).48h after transfection,QPCR was used to confirm the interfering efficacy to ensure that siRNA can be used for following experiments.CCK-8 assay,EdU cell immunofluorescence staining assay,Hoechst 33342 staining assay,wound healing assay were used to detect the cytoactive,proliferation,apoptosis and migration of SKOV3 cells after transfection.The related protein of PTEN/Akt signaling pathway was evaluated by Westem blotting.Results The level of mGluR5 mRNA in transfection group was (18.3 ± 2.3) ％,significantly decreased compared with NC (P＜0.05).72h after transfection,the absorbance of transfection group was 2.4±0.3,lower than 4.1±0.2 of NC (P＜0.05).48h after transfection,the proliferative rate of SKOV3 cells was (9.2± 1.2)％ in transfection group,lower than (22.4±2.3) ％ of NC (P＜0.05);the cell apoptotic rate of transfection group was (28.2±2.2) ％,higher than (6.2± 1.3) ％ of NC (P＜0.05).In wound healing assay,relative wound closure rates of NC and transfection group were (100.0±2.1)％ and (48.6±4.3)％ (P＜0.05).Western blotting showed that p-Akt expression decreased,and PTEN expression increased after transfecion (P＜0.05),but the expression of Akt did not change obviously (P＞0.05).Conclusion Silencing the expression of mGluR5 can inhibit the proliferation,migration and enhance apoptosis of SKOV3 cells.PTEN/Akt signaling pathway may be involved in this process.