| Backgrounds and Objectives Multiple myeloma(MM)is the second most common hematologic malignancy after non-Hodgkin's lymphoma and accounts for 13% of malignant hematologic diseases.There is no cure yet.Bortezomib(BTZ)is the first generation of proteasome inhibitor,its efficacy on multiple myeloma is significant,PD-based program is the first-line treatment recommended by domestic and foreign guidelines,and bortezomib)Entered China's National Health Insurance Catalog in September 2017,allowing more Chinese patients with multiple myeloma to use this medicine.However,in order to further improve the curative effect,the addition of adjuvant drugs on the basis of PD is still a challenge that hematologists face when treating MM.Recombinant mutant of human TRAIL(rmh TRAIL)is a recombinant protein antitumor drug that was developed by Beijing Shadong Biotechnology Co.,Ltd.and is undergoing rmh TRAIL-based regimen.Clinical trials for relapsed multiple myeloma.rmh TRAIL combined with bortezomib may be more suitable for Chinese patients with multiple myeloma.Epigenetics has a major guiding role in the stratification,treatment,and medication of diseases.Micro RNAs can affect life activities in many aspects such as regulation of transcription,expression of proteins,and the like.mi R-17-5p and mi R-886-5p are two important micro RNAs that regulate tumor expression.They have been widely studied in breast cancer,gastric cancer,endometrial cancer,and other malignancies.There is no single and multiple myeloma.Related reports.From the background,this topic aims to explore the following issues.1.To explore the effect of rmh TRAIL monotherapy on the proliferation inhibition of multiple myeloma cell lines.2.Changes in mi R-17-5p and mi R-886-5p in multiple myeloma after rmh TRAIL drug treatment.3.To explore the effect of BTZ monotherapy and rmh TRAIL combined with BTZ on proliferation inhibition of multiple myeloma cell lines.4.Changes in mi R-17-5p and mi R-886-5p in multiple myeloma treated with BTZ alone and Rmh TRAIL combined with BTZ.Methods 1.Human multiple myeloma cell lines U266B1 and NCI-H929 were cultured.The effect of rmh TRAIL(0/78.125/312.5/1250/5000/20000 ng/ml)on the inhibition of proliferation was measured by CCK-8 method,and apoptotic rate was measured by flow cytometry.MM cells were treated with drugs and the mi Rs were extracted.-886-5p and mi R-17-5p,comparing the differences in the content,through a statistical analysis of the correlation with drug effects.2.Human multiple myeloma cell lines U266B1 and NCI-H929 were cultured.The effect of rmh TRAIL(20000ng/ml)combined with different concentrations of BTZ(10/20/40/80/160 n M)on the proliferation inhibition was measured by CCK-8 method,and the apoptosis rate was measured by flow cytometry.After MM cells were treated with drugs,mi R-886-5p and mi R-17-5p were extracted,and the difference in the content was compared.Statistical analysis was performed to show whether the drug was related to the drug effect.Results 1 The effect of rmh TRAIL on proliferation inhibition of multiple myeloma cells detected by CCK-8 assay.rmh TRAIL has a proliferation-inhibitory effect on multiple myeloma NCI-H929 cell lines.Different doses of rmh TRAIL(0/78.125/312.5/1250/5000/20000 ng/ml)RMHTRAIL acted on NCI-H929 cells,and the trend of proliferation inhibition rate at 24 h was(-1.34±0.45)% increased to(35.66±0.39)%.The inhibitory rate of proliferation increased from(6.53±0.33)% to(52.23±0.96)% at 48 h,and increased from(19.73±0.84)% to(70.04±0.56)% at 72 h,in a timedependent and concentration-dependent manner.P<0.05).The IC50 values of RMHTRAIL-treated multiple myeloma cells after 24 h,48 h,and 72 h were(63579.532±4.803)ng/ml,(15580.413±4.193)ng/ml,(5181.069±3.714)ng/ml,respectively.In sharp contrast with NCI-H929 cells,different doses of rmh TRAIL(0/78.125/312.5/1250/5000/20000ng/ml)rmh TRAIL acted on U266B1 cells,and the trend of proliferation inhibition rate at 24 h was(-5.62±0.24).)% increased to(4.66±0.14)%,48 h proliferation inhibition rate increased from(2.23±0.32)% to(7.84±0.36)%,72 h proliferation inhibition rate increased from(1.73±0.32)% to(9.04±1.26)%,does not show time-dependent and concentration-dependent(P>0.05).2 Annexin V-FITC/PI Double Staining Method for Detection of Apoptosis Rate after RMHTRAIL Treatment of Multiple Myeloma After taking RMHTRAIL IC50 concentration for 48 h in U266B1 and NCI-H929 cells,the apoptosis rate of U266B1 cells increased from(6.61±0.67)% to(7.06±0.28)%,while the apoptosis rate of NCI-H929 cells was(5.59).±0.03% increase to(27.13±1.35)%.Using SPSS software for statistical analysis,it was found that RMHTRAIL significantly inhibited the proliferation of NCI-H929(p<0.05),but there was no statistical significance for U266B1 cells(p>0.05).3 Effect of rmh TRAIL on mi R-17-5p and mi R-886-5p in MM CellsAfter rmh TRAIL treatment of U266B1 cells,the changes of mi R-17-5p and mi R-886-5p in the cells were not statistically significant.After RMHTRAIL treatment of NCI-H929 cells,the changes in the levels of mi R-17-5p and mi R-886-5p were statistically significant,and the contents of both were significantly reduced.4 CCK-8 assay for the effect of BTZ and rmh TRAIL combined with BTZ on proliferation inhibition of multiple myeloma cells rmh TRAIL has a proliferation-inhibitory effect on multiple myeloma NCI-H929 cell lines.Different doses of BTZ(10/20/40/80/160 n M)acted on NCI-H929 cells,and the trend of proliferation inhibition rate at 24 h was(6.79±0.79)% increased to(42.02±0.56)%,48 h inhibition of proliferation from(12.14±0.85)% increased to(69.99±0.73)%,and 72 h proliferation inhibition rate increased from(14.10±0.51)% to(81.70±1.28)% in a time-dependent and concentrationdependent manner(P<0.05).The IC50 values of BMSCs treated with multiple myeloma cells 24 h,48 h,and 72 h were(211.95±1.87)ng/ml,(73.78±1.87)ng/ml,and(56.22±1.75)ng/ml,respectively.Different doses of BTZ(10/20/40/80/160 n M)acted on U266B1 cells,and the trend of proliferation inhibition rate at 24 h was(6.29±0.31)% increased to(9.25±0.59)%,and the inhibition rate of 48 h proliferation was(7.54).±0.21)% increased to(56.97±0.85)%,and the 72 h proliferation inhibition rate increased from(12.71±0.22)% to(70.23±0.76)%,showing no time-dependent and concentration-dependent(P<0.05).The IC50 values after 24 h,48 h,and 72 h of BTL-treated multiple myeloma U266B1 lines were(216.13±2.33)ng/ml,(121.68±2.09)ng/ml,and(72.03±1.86)ng/ml,respectively.RMHTRAIL combined with BTZ resulted in similar results to BTZ alone.Different doses of BTZ(10/20/40/80/160 n M)acted on U266B1 cells,and the proliferation inhibition rate at 24 h was(2.61±0.90)% increased to(54.26±0.72)%,and the inhibition rate of 48 h proliferation was(8.73).(±0.65)% increased to(53.95±1.46)%,and the 72 h proliferation inhibition rate increased from(11.63±0.85)% to(72.88±1.24)% in a time-dependent and concentrationdependent manner(P<0.05).RMHTRAIL was combined with different doses of BTZ(10/20/40/80/160 n M)in NCI-H929 cells.The trend of proliferation inhibition rate at 24 h was(55.91±0.48)% increased to(92.56±0.54)%,48 h inhibition of proliferation.The rate increased from(60.25±1.30)% to(92.58±1.52)%,and the 72 h proliferation inhibition rate increased from(72.52±0.81)% to(93.23±1.09)%,showing no time-dependent and concentration-dependent(P<0.05).).5 Annexin V-FITC/PI Double Staining Method for Detecting Apoptosis Rate of rmh TRAIL in Treating Multiple Myeloma The apoptosis rate of U266B1 cells increased from(5.89±0.18)% to(26.43±0.86)% after taking IC50 concentration of BTZ for 48 h in U266B1 and NCI-H929 cells,and the proliferation inhibition rate of double drugs was(26.95±1.87).).The apoptosis rate of NCI-H929 cells increased from 8.78±0.99)% to(24.32±0.53)%,and the proliferation inhibition rate of the two drugs combined reached(30.35±1.35).Using SPSS software for statistical analysis,it was found that the inhibition of proliferation of NCI-H929 and NCIH929 cells by BTZ group and RMHTRAIL combined with BTZ group was statistically significant(p<0.05).6 Effects of BTZ and rmh TRAIL Combined with BTZ on mi R-17-5p and mi R-886-5p in MM Cells There was no statistically significant change in the content of mi R-17-5p and mi R-886-5p in cells treated with BTZ alone and RMHTRAIL combined with BTZ.After treatment with BTZ monotherapy and RMHTRAIL combined with BTZ,the changes of mi R-17-5p and mi R-886-5p content were statistically significant,and the contents of both were significantly reduced.Summaries 1.rmh TRAIL has a growth inhibitory effect on multiple myeloma cell line NCIH929,and has no proliferation inhibiting effect on multiple myeloma cells U266B1.2.rmh TRAIL treatment of NCI-H929 cells,in which the reduction of mi R-17-5p and mi R-886-5p was statistically significant,but was not statistically significant for U266B1 cells.3.BTZ has obvious inhibitory effect on MM cell U266B1 and NCI-H929.BTZ combined with rmh TRAIL has a significant inhibitory effect on NCI-H929,and the two drugs show a synergistic effect,and also have an inhibitory effect on proliferation of U266B1 cells,but the two drugs have no synergistic effect.4.rmh TRAIL combined with BTZ treatment of multiple myeloma cells NCI-H929 and U266B1,mi R-17-5p and mi R-886-5p reduction was statistically significant. |
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