The Expression Of Robo1 In Cervical Cancer And Its Effect On Cervical Cancer Cells’ Proliferation,Migration And Invasion

Objective:To investigate the expression of Slit2/Robo1 in human cervical cancer tissues and the relationship between Robo1 and clinicopathology,and to analyze the effect on cervical cancer cells’ proliferation,migration and invasion.Methods:(1)A total of 70 cervical cancer tissues were collected from patients with cervical cancer who received surgical treatment in the department of gynaecology of People’s Hospital of Guangxi Zhuang Autonomous Region from December 2013 to December 2019,and 20 normal cervical tissues from benign uterine lesions(such as fibroids,adenomyosis,etc.)were selected as the control group.Quantitative real time polymerase chain reaction(q RT-PCR)was used to detect the expression of Slit2/Robo1 m RNA in cervical tissues,and to analyze the relationship between the expression of Robo1 and clinicopathological parameters;(2)Overexpressing Robo1 lentiviral vector with Puromycin resistance gene and green fluorescent protein(GFP)was constructed by lentiviral vector technology.Cervical cancer SIHA and He La cells with stable overexpressing Robo1 were screened by Puromycin.(3)The cell proliferation was detected by clonal formation assay and CCK8 assay.The migration ability of cells was detected by scratch assay and transwell cell migration assay.Transwell cell invasion assay was used to detect cell invasion ability.Results:(1)The relative expression level of Robo1 m RNA in 70 cervical cancer tissue samples was lower than that in 20 normal cervical tissue samples(P<0.05).The low expression of Robo1 m RNA was associated with low differentiation of tumor(P<0.05),late FIGO stage(P=0.003)and lymph node metastasis(P<0.05);(2)Cervical cancer stable Si Ha and He La cells expressing Robo1 were screened.MTT and clone formation experiments showed that the proliferation ability of Siha and He La cells in the over-expression group(OE)was lower than that in the NC group,and the difference was statistically significant(P<0.05).Compared with the control group,the inhibitory effect of Robo1 on the growth of He La and Siha cells in the experimental group was the most obvious on the 5th day(P value was 0.0004 and 0.002,respectively).The scratch healing rate of He La and Si HA cells in OE group was lower than that in NC group(P< 0.05).The number of cells in the OE group of He La and Siha cells penetrating the basement membrane was less than that in the NC group(P< 0.05).Transwell invasion assay showed that the number of cells penetrating matrix membrane in the OE group of He La and Siha cells was less than that in the NC group(P<0.05).Conclusion:(1)The expression of SLIT2 /Robo1 was low in cervical cancer tissues.(2)To observe the expression of Robo1 m RNA in cancer tissue is helpful to determine the degree of differentiation,FIGO stage and lymphatic metastasis of cervical cancer;(3)Robo1 can inhibit the proliferation,migration and invasion of cervical cancer,and it may be involved in the occurrence and development of cervical cancer as a tumor suppressor gene.