Objective To analyze the expression of circRNA0087432 (hsa_circ_0087432) in serum exosomes of patients with cervical cancer, and explore its possible relationship with cervical cancer metastasis. Methods The cervical cancer tissue and serum of 30 patients, who were diagnosed as cervical squamous cell carcinoma from June 2019 to June 2020 in Guangzhou Panyu District Central Hospital and untreated with radiotherapy and chemotherapy before surgery, were collected as the experimental group.Normal cervical tissue and serum of 30 women without cervical cancer were used as control group. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression level of hsa_circ_0087432 in cervical tissues and serum exosomes of patients with cervical cancer and healthy people; Siha cells were transfected with plasmid to overexpress hsa_circ_0087432. According to different treatments, Siha cells were divided into three groups: control group (untransfected plasmid), vector group (transfected with pLC5-ciR) and hsa_circ_0087432 group (transfected with hsa_circ_0087432-pLC5-ciR). The exosomes were collected and extracted, and the exosyndrome were characterized by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA)and Western blotting; Endothelial cells were divided into three groups according to different treatments: control-Exs group (extracted exosomes from control group), vector-Exs group (extracted exosomes from vector group) and hsa_circ_0087432-Exs group (extracted exosomes from hsa_circ_0087432 group). CCK-8 kit was used to detect the effect of exosomes on proliferation of endothelial cells.The effect of exosomes on the migration of endothelial cells were detected by scratch test and Transwell. Results The results of TEM showed that most of the exosomes of Siha cells were elliptical or round. The particle size analysis showed that their diameters ranged of 30-150 nm. Western blotting showed that the expressions of CD9, CD81 and CD63 in the two groups of vesicles were positive. The qPCR results showed that the expression level of hsa_circ_0087432 in cervical tissue was significantly higher in experimental group than that in control group (4.03±1.51 vs. 1.00±0.26, P<0.01), and the expression level of hsa_circ_0087432 in serum exosomes was also obviously higher in experimental group than that in control group (1.97±0.04 vs. 1.02±0.23, P<0.01); CCK-8 results showed that, compared with the control-Exs group, the proliferation of human umbilical vein endothelial cells was significantly promoted in hsa_circ_0087432-Exs group (1.57±0.04 vs. 1.09±0.11, P<0.05). Scratch experiments showed that, 12 hours after treatment, the healing degree was better in hsa_circ_0087432-Exs group than that in control-Exs group (24.66%±2.92% vs. 15.01%±3.12%, P<0.05); and 24 hours after scratching, the healing degree was even better in hsa_circ_0087432-Exs group than that in control-Exs group (74.84%±13.22% vs. 38.70%±2.12%, P<0.01). Transwell also showed that the counted number of endothelial cells increased obviously in hsa_circ_0087432-Exs group than that in vector-Exs group and control-Exs group (1755.00±97.35 vs. 1218.00±103.53 vs. 1044.00±103.79) with significant difference (P<0.05). Conclusion The hsa_circRNA 0087432 is highly expressed in serum exosomes of patients with cervical cancer, and the exosomes derived from cervical cancer cells which over-pressing the hsa_circ_0087432 can promote the proliferation and migration of human umbilical vein endothelial cells.
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |