High Throughput Sequencing Technology Was Used To Screen MiRNA In Ovarian Cancer Nude Mouse Models Treated With Zedoary Turmeric Oil

Objective:The aim of this study was to determine miRNA expression level in human ovarian cancer nude mouse tissues after treatment with Zedoary turmeric oil by high-throughput sequencing technology,and to screen out specific miRNA indicators and analyze them.Methods:Fifty female BALB/c nude mice aged 5 to 6 weeks at 18 to 20g were selected.42 nude mice were randomly inoculated with 0.2mL 2×107 ·mL-1 human ovarian cancer SKOV3 cell suspension subcutaneously under the right axilla.The remaining 8 nude mice were subcutaneously injected 0.2mL rpmi-1640 medium under the right axilla as the blank group（30ml·kg-1（normal saline））.Using vernier calipers,tumor diameter was measured in nude mice injected with human ovarian cancer SKOV3 cell suspension,The nude mice were randomly divided into model group（30ml·kg-1（normal saline））,zedoary turmeric alcohol group（10mg·kg-1）,paclitaxel group（5mg·kg-1）,zedoary turmeric oil high-dose group（200mg·kg-1（zedoary turmeric oil injection）and zedoary turmeric oil medium-dose group（100mg·kg）-1（Zedoary turmeric oil injection）,zedoary turmeric oil low-dose group（50mg·kg-1（Zedoary turmeric oil injection））;A total of 7 groups,7 in each group（8 in blank group）,a total of 50;Intraperitoneal injection（INTRaperitoneal injection）was given once every other day for a total of 14 times for 28 consecutive days.DNBSEQ,Agilent 2100 and Bowtie2 were used for results determination,data collation and data analysis.Results:（NCBI database and CNKI were used for results analysis;Filter:|log2FC|1＆Qvalue acuities were 0.001 or higher）;（1）Compared with model group,11 mirnas were differentially expressed in zedoary turmeric oil high-dose group,including 6 mirnas related to ovarian cancer,including 3 up-regulated mirnas:HSA-Mir-199a-3p,HSA-Mir-205-3p,and HSA-Mir-205-5p.Three down-regulated genes were hS A-Mir-133 A-3 p,HSA-Mir-133b and HSA-Mir-423-5p,respectively.（2）One miRNA was differentially expressed in zedoary turmeric oil high dose group and medium dose group,among which 0 miRNA was related to ovarian cancer.（3）Compared with the zedoary turmeric oil high-dose group and the zedoary turmeric oil low-dose group,9 mirnas were differtively expressed,including 3 mirnas related to ovarian cancer:2 up-regulated mirnas were hSA-Mir-206 and HSA-Mir-512-3p,respectively.One was down-regulated as HSA-Mir-423-5p;（4）Compared with the zedoary turmeric oil high-dose group and zedoary Turmeric alcohol group,10 mirnas were differentially expressed,among which 4 mirnas related to ovarian cancer were upregulated,3 were hSA-Mir-199a-3p,HSA-Mir-205-3p,and HSA-Mir-205-5p,respectively.One was down-regulated as HSA-Mir-423-5p;Compared with the paclitaxel group,there were 4 differentially expressed mirnas in the high-dose zedoary turmeric oil group,including 3 mirnas related to ovarian cancer,including 2 up-regulated mirnas（hSA-Mir-205-3p and HSA-Mir-205-5p）.One was down-regulated as HSA-Mir-423-5p;6 mirnas were differentially expressed in zedoary turmeric oil medium-dose group and model group,among which 4 mirnas were related to ovarian cancer,0 were up-regulated.Four down-regulated genes were hSA-Mir-133A-3p,HSA-Mir-133b,HSA-Mir-206 and HSA-Mir-423-5p,respectively.There were 6 differentially expressed mirnas in zedoary zedoary oil medium-dose group and low-dose group,including 1 miRNA related to ovarian cancer,0 miRNA up-regulated.One was down-regulated as HSA-Mir-423-5p;By comparing the medium-dose zedoary turmeric oil group with the Zedoary Turmeric alcohol group,there were 3 mirnas with different expression,of which 2 mirnas were associated with ovarian cancer（0 up-regulated）.Two down-regulated genes were hS A-Mir-133 A-3 p and HSA-Mir-133b.Compared with paclitaxel group,there were 3 differentially expressed mirnas in zedoary turmeric oil medium-dose group,among which 1 miRNA was related to ovarian cancer,0 miRNA was up-regulated.One was down-regulated as HSA-Mir-423-5p;Compared with the model group,24 mirnas were differtively expressed in the low-dose zedoary oil group,including 6 mirnas related to ovarian cancer:2 up-regulated mirnas:HSA-Mir-1246 and HSA-Mir-199a-3p.Four were down-regulated,namely,HS A-Mir-133 a-3 p,HSA-Mir-133b,HSA-Mir-206,and HSA-Mir-512-3p.11 Zedoary turmeric oil low dose group and zedoary turmeric alcohol group were compared,there were 14 differentially expressed mirnas,including 4 mirnas related to ovarian cancer,0 were up-regulated;Four were down-regulated,namely,HSA-Mir-133 a-3 p,HSA-Mir-133b,HSA-Mir-206,and HSA-Mir-512-3p.（12）Differentially expressed miRNA in zedoary turmeric oil low-dose group and paclitaxel group,including 1 miRNA related to ovarian cancer,0 miRNA up-regulated;One was down-regulated as HSA-Mir-512-3p;Compared with model group,5 mirnas were differentially expressed,among which 0 mirNAS were related to ovarian cancer.Eight mirnas（14）were differentially expressed in zedoary alcohol group and paclitaxel group,among which four mirnas（3）were up-regulated（HSA-Mir-133A-3p,HSA-Mir-133b,hSA-Mir-206）related to ovarian cancer.One was down-regulated as HSA-Mir-199a-3p;Compared with participant paclitaxel group,13 mirnas were differentially expressed,including 4 mirnas related to ovarian cancer.One miRNA was up-regulated（HSA-Mir-199a-3p）.Three down-regulated genes were hSA-Mir-133A-3p,HSA-Mir-133b and HSA-Mir-206,respectively.（16）The blank group did not form tumor tissue,so it was not involved in the comparison.Three mirnas were expressed in the high-dose,medium-dose and low-dose groups of Zedoary oil,among which two mirnas related to ovarian cancer were down-regulated,namely,HSA-Mir-133A-3p and HSA-Mir-133b.Zedoary turmeric oil in high dose group,zedoary turmeric oil dose group,zedoary turmeric oil two comparison between the low dose group were not significant differences in expression of three groups were compared with the model group had a significant difference expression（filter condition:|log2FC| 1＆Qvalue acuities were 0.001 or higher）;The results of GO functional enrichment analysis and KEGG pathway enrichment analysis were as follows:A total of 574 items were obtained by GO functional enrichment analysis（screening condition:P value≤0.05）,including 351 items for Biological process;Cellular Component has 94 entries;Molecular Function has 129 entries;Enrichment analysis of KEGG pathway（Screening conditions:P value≤0.05）.By reviewing their pathway annotation and disease annotation information,it was found that HSA-Mir-133A-3p and HSA-Mir-133b play a role in ovarian cancer mainly by targeting PIK3CD to regulate phosphoinositol metabolism.Conclusion:1）Two mirnas co-expressed in zedoary turmeric oil high-dose group,zedoary turmeric oil medium-dose group and zedoary turmeric oil low-dose group were down-regulated,respectively,as HSA-Mir-133A-3p.Hsa-miR-133 b;2)On hSA-Mir-133A-3p;In terms of the expression of HSA-Mir-133b in each sample,there was no dose-effect relationship between zedoary turmeric oil and ovarian cancer treatment in this study.3)KEGG pathway enrichment analysis results showed that HSA-Mir-133A-3p and HSA-Mir-133b play a role in ovarian cancer mainly by targeting PIK3CD to regulate phosphoinositol metabolism.