Effect Of Non-platelet RNA-containing Particle-derived Cells From Umbilical Cord Blood On Lung Cancer A549 Cells

Objective:To study the effects of non-platelet RNA-containing particles(NPRCP)on the proliferation,cell cycle,and surface markers of lung cancer cell line,different concentrations of NPRCP from umbilical cord blood were co-cultured with the cell line.Method:(1)NPRCP was isolated from the cord blood supernatant.Different concentrations of NPRCP were co-cultured with A549 cells for 24h,48h,72h;and then the MTT method was used to detect the inhibitory rate of different concentrations of NPRCP on A549 cell growth.(2)Different concentrations of NPRCP were co-cultured with A549 cells,and the effects of different concentrations of NPRCP on surface markers and cell cycle of A549 cells were measured by flow cytometry after co-culture for 48h.Results:(1)After co-culturing NPRCP with A549cells for 24h,the inhibition rate of A549 cell was(0.01±0.07)%in group A(NPRCP concentration was 0.5*10~5/?l),(0.02±0.05)%in group B(NPRCP concentration was 1*10~5/?l),(0.04±0.11)%in group C(NPRCP concentration was 2*10~5/?l),(0.06±0.10)%in group D(NPRCP concentration was 4*10~5/?l),(0.13±0.08)%in group E(NPRCP concentration was 8*10~5/?l).After co-culturing NPRCP with A549 cells for 48h,the inhibition rate of A549cell was(0.01±0.07)%in group A,(0.03±0.15)%in group B,(0.06±0.13)%in group C,(0.12±0.10)%in group D,(0.13±0.05)%in group E.After co-culturing NPRCP with A549cells for 72h,the inhibition rate of A549 cell was(0.01±0.08)%in group A,(0.08±0.13)%in group B,(0.12±0.13)%in group C,(0.14±0.16)%in group D,(0.16±0.20)%in group E.After co-culturing NPRCP with A549 cells for 24h,48h,and 72h,the three groups of data showed that as the concentration of NPRCP increased,the inhibition rate of A549 cell growth gradually increased(P<0.05).(2)After co-culturing NPRCP and A549 cells for 48h,the cell cycle of A549 cells in each group was measured by flow cytometry.The ratio of G0/G1 phase was(64.77±2.32)%in the control group(NPRCP concentration was 0/?l),(65.82±1.61)%in group A(NPRCP concentration was 0.5*10~5/?l),(66.11±2.13)%in group B(NPRCP concentration was 1*10~5/?l),(63.84±0.47)%in group C(NPRCP concentration was2*10~5/?l),(67.25±1.21)%in group D(NPRCP concentration was 4*10~5/?l),(65.39±3.72)%in group E(NPRCP concentration was 8*10~5/?l).There was no significant difference between the experimental(A-E group)and the control groups(P>0.05).There was no significant difference between the experimental groups(P>0.05).The ratio of S phase in control group,group A,group B,group C,group D,group E were:(29.76±2.41)%,(28.92±0.52)%,(27.98±2.07)%,(30.78±0.81)%,(27.88±0.41)%,(28.44±3.17)%.There was no significant difference between the experimental(A-E group)and the control groups(P>0.05).There was no significant difference between the experimental groups(P>0.05).The ratio of G2/M phase in control group,group A,group B,group C,group D,group E were:(5.47±0.4)%,(6.45±0.22)%,(5.91±0.85)%,(5.7±0.99)%,(5.87±1.02)%,(6.17±1.51)%.There was no significant difference between the experimental(A-E group)and the control groups(P>0.05).There was no significant difference between the experimental groups(P>0.05).After co-cultivating different concentrations of NPRCP with A549 cells for 48h,NPRCP had no effect on the cell cycle distribution of A549 cells(P>0.05).Calculate the proliferation index(PI)of each group of cells according to the formula.The PI of control group was(0.35±0.02)%,group A was(0.34±0.02)%,group B was(0.34±0.02)%,group C was(0.37±0.01)%,group D was(0.33±0.01)%,group E was(0.35±0.04)%.There was no significant difference between the experimental(A-E group)and the control groups(P>0.05).There was no significant difference between the experimental groups(P>0.05).(3)The expression rates of cell surface marker CD133 in the control and experimental groups(groups A,B,C,D,E)were:(10.14±2.16)%?(10.10±3.47)%?(10.48±2.83)%?(10.32±2.89)%?(10.53±2.47)%?(10.64±2.90)%.There was no significant difference in the expression rate between the experimental and the control groups and between the experimental groups(P>0.05).The expression rates of cell surface marker CD44 in the control and experimental groups were:(99.06±0.09)%?(99.05±0.21)%?(99.09±0.04)%?(98.95±0.06)%?(99.11±0.04)%?(98.98±0.06)%.There was no significant difference between the experimental and the control groups(P>0.05).There was no significant difference between the experimental groups(P>0.05).The expression rates of cell surface marker CD24 in the control and experimental groups were:(10.29±2.78)%?(9.88±3.85)%?(8.92±2.73)%?(9.13±3.26)%?(10.12±3.01)%?(9.28±2.86)%respectively.There was no significant difference between the experimental and the control groups(P>0.05).In addition there were no significant difference between the experimental groups(P>0.05).The expression rates of the cell surface marker CD338(ABCG2)in the control group and experimental groups were(15.26±4.23)%?(16.22±2.27)%?(15.12±5.08)%?(15.39±2.79)%?(13.34±2.98)%?(10.81±1.80)%.The difference in CD338 expression rate between the experimental and the control groups was not significant(P>0.05).The comparison between the experimental groups was not significant(P>0.05).Conclusions:(1)With the increase of NPRCP concentration,the inhibition rate of A549 cells increased gradually.(2)Different concentrations of NPRCP had no effect on the cell cycle of A549 cells,and had no effects on the proliferation index of A549 cells.(3)NPRCP at different concentrations had no effects on the expression of CD133,CD338,CD24 and CD44 of A549 cells.