| Background: Cervical cancer is the third most common cancer among women in the world and the second most common cancer among women in China.It is also the leading cause of gynecological tumor-related death worldwide and seriously endangers women’s health.At present,concurrent chemoradiotherapy and brachytherapy are important components of the standard treatment strategies for cervical cancer,but there are still some patients who fail treatment due to radiotherapy resistance.Long non-coding RNA（lncRNA）is a kind of RNA with transcription length over200 bp,which is the main transcription product in human but incapable of encoding proteins.Many studies have also shown that lncRNA plays important roles in many biological process including gene expression,transcription and cell proliferation through m RNA shearing,epigenetic silencing and lncRNA-mi RNA interaction.A number of studies have shown that lncRNA plays an important role in the occurrence,development and prognosis of cervical cancer.Moreover,the mechanism of chemoradiotherapy resistance and tumor recurrence has become the focus of cervical cancer research.Objective: We conducted whole exome sequencing on RNA extracted from tissues of 26 patients with locally advanced cervical cancer in previous research,and survival analysis show that 7 differential genes were associated with RFS of patients.To validate the gene expression,we conduct q RT-PCR in independent samples.Subsequently,in vitro experiments were performed to explore the correlation between the gene and radiosensitivity of cervical cancer,and its potential mechanism.Our study aims to probe new targets for predicting the effect of radiotherapy,and provide new strategies for improving the survival rate and prognosis.Methods: 66 patients with cervical cancer（FIGO stage IIb-IVa）were enrolled into study who underwent radical chemoradiotherapy from 2015 to 2017,divided them into early recurrence（radiotherapy resistant）and late recurrence group（radiotherapy sensitive）,and q RT-PCR was used to validate the expression of differential genes.Human cervical cancer cell lines He La and Ca Ski were selected to compare the differential gene expression by q RT-PCR.The target genes were up/down regulate through lentivirus transfection.Clony formation assay,scratch assay,western blot assay,cell cycle and apoptosis assay were performed to investigate the relationship between gene and radiotherapy sensitivity of cervical cancer cells.Results:（1）The expression of CTD-2292M14.1 was significantly increased in early recurrence（ER）group from 66 independent samples（p <0.05）.The expression of CTD-2292M14.1 gene in He La cells was higher than that in Ca Ski cells（p <0.05）.He La cells were selected to cultivate the permanent lentivirus transfection cell line.（2）The expression of CTD-2292M14.1 in LV-CTD group was higher than that in control group（p <0.05）,the survival fraction increased after radiation treatment（p<0.05）,and the expression of CTD-2292M14.1 in LV-CTD-RNAi group was lower than that in control group（p <0.05）.The survival fraction decreased after radiation treatment（p <0.05）.These results suggest that the overexpression of CTD-2292M14.1reduces the sensitivity of cervical cancer cells to radiotherapy.（3）The scratch healing and proliferation rate of irradiated LV-CTD group were significantly increased compared with control group（p <0.05）,while the scratch healing rate and proliferation of LV-CTD-RNAi group were decreased compared with control group（p <0.05）.Overexpression of CTD-2292M14.1 enhanced migration and proliferation of cervical cancer cells after irradiation.（4）Flow cytometry showed that the overexpression of CTD-2292M14.1 could not inhibit the apoptosis of cervical cancer cells after irradiation.（5）Cell cycle analysis presented that LV-CTD+8Gy group had a significantly increased cells in proportion of S phase compared with control+8Gy group（p <0.05）,overexpression of CTD2292M14.1 arrested more He La cells in S phase.LV-CTD-RNAi+8Gy had a significant increase in proportion of G1 phase compared with control+8Gy group（p <0.05）,indicating G1 phase arrest.Overexpression of CTD-2292M14.1 promoted the cycle progression of cervical cancer cells after irradiation.（6）There was no significant correlation between LC3 and P62 expressions in all groups.After irradiation,the expression of p-AMPK/AMPK and p-LKB1/LKB1 in LV-CTD+8Gy group were higher than LV-CTD group（p < 0.05）.The results showed that CTD2292M14.1 regulates the radiosensitivity of cervical cancer through AMPK/LKB1 pathway.Conclusions: CTD2292M14.1 may affect the proliferation,migration and cell cycle of cervical cancer through AMPK/LKB1 pathway,thus regulating the radiosensitivity of cervical cancer. |
PDF Full Text Request