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1.
Chinese Journal of Oncology ; (12): 742-747, 2019.
Artículo en Chino | WPRIM | ID: wpr-796929

RESUMEN

Objective@#To investigate the effect of thymopoietin (TMPO) gene deleted by small interfering RNA (RNAi) on the proliferation and apoptosis of lung cancer cell A549 and its mechanism.@*Methods@#TMPO siRNA was transfected into A549 cells by lipofection. The transfected siRNA control was used as a negative control, and the parent cells were used as blank control. Forty-eight hours later, the expression of TMPO in the transfected cells was detected by real-time fluorescent quantitative polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay, cell cycle and apoptosis were detected by flow cytometry, the protein levels of proliferating cell nuclear antigen (PCNA), cleaved caspase-3, notch receptor 1 (Notch1) and mammalian rapamycin target protein (mTOR) were detected by Western blot analysis.@*Results@#The results of RT-PCR and Western blot showed that the expression levels of TMPO mRNA in the blank control group, the negative control group and TMPO siRNA transfected group were (1.01±0.11), (0.99±0.10), (0.36±0.04), respectively, the protein levels were (0.27±0.02), (0.29±0.03), (0.08±0.10), respectively. The expression levels of TMPO mRNA and protein in the transfected group were significantly lower than those in the blank control and negative control group (P<0.05). The results of MTT assay showed that the OD values of the blank control group, the negative control group and the transfected group were (0.35±0.04), (0.37±0.04) and (0.34±0.03) at 24 h of transfection, respectively. The OD values at 48 h were (0.47±0.06), (0.46±0.08), (0.37±0.04), the OD values at 72 h were (0.75±0.08), (0.73±0.07), (0.49±0.05), respectively, and the OD values at 96 h were (1.09±0.07), (1.06±0.08), (0.56±0.06). The proliferation abilities of the transfected cells at 48, 72, 96 h were significantly lower than those of the blank control and the negative control group (P<0.05). The results of flow cytometry showed that the proportion of G0/G1 phase cells in blank control group, negative control group and transfection group were (62.55±2.03)%, (61.24±3.15)%, (47.35±2.44)%, respectively. The proportion of cells in S phase were (17.12±1.31)%, (17.70±2.01)%, and (20.81±2.06)%, respectively. The proportion of cells in G2/M phase were (20.33±1.43)%, (21.06±1.52)%, (31.84±2.76)%, respectively. The proportion of cells in G0/G1 phase of transfection group was significantly lower than those of blank control and negative control group (P<0.05). The proportion of cells in G2/M phase of transfection group was significantly higher than those of blank control and negative control group (P<0.05). The apoptosis ratio of the transfection group was (34.10±2.69)%, significantly higher than (2.96±0.03)% of the blank control and (3.01±0.04)% of the negative control group (P<0.05). Western blot analysis showed that PCNA, Notch1 and mTOR proteins were down-regulated while cleaved caspase-3 protein was up-regulated in A549 cells after deletion of TMPO.@*Conclusion@#The inhibition of TMPO gene expression induced by small interfering RNA can significantly inhibit the proliferation and induce apoptosis of A549 cells, and the mechanism is associated with the inhibition of the activation of Notch1/mTOR signaling pathway.

2.
Chinese Journal of Primary Medicine and Pharmacy ; (12): 3179-3183, 2017.
Artículo en Chino | WPRIM | ID: wpr-614675

RESUMEN

Objective To investigate the effect of thymopentin combined with routine medication on serum interleukin 17A(IL-17A),chemokine 12 (CXCL12) levels and transient receptor potential channel 1 (TRPC1)expression in elderly patients with chronic obstructive pulmonary disease (COPD).Methods 156 COPD patients were selected as study objects.They were randomly divided into two groups(n =78) according to the digital table.The control group received routine treatment,and the observation group received subcutaneous injection of thymopentin on the basis of routine treatment.Before and after treatment,the CD3+,CD4+,CD8+,CD4+/CD8+ levels were tested to compare the immunologic function of the two groups.Before and after treatment,the FEV1,FEV1/FVC,PEF were tested to compare the pulmonary ventilation function of the two groups.The IL-17A,CXCL12 levels in peripheral blood and TRPC1 level in bronchoalveolar liquid before and after treatment were tested and compared between the two groups.The adverse effect was compared.Results Before treatment,the CD3+,CD4+,CD8+ and CD4+/CD8+ between the two groups had not statistically significant differences (all P > 0.05),which of the control group after treatment had no significant change (all P > 0.05),the CD3+,CD4+,CD4+/CD8+ of the observation group [(65.17 ± 2.39) %,(41.06 ±2.15) %,(1.50 ± 0.74) %] were significantly higher than control group [(52.66 ± 2.38) %,(32.30 ± 2.05) %,(0.80 ± 0.81) %] (all P < 0.05),and the CDs+ of the observation group [(24.02 ± 2.23) %] was significantly lower than control group[(32.66 ± 1.97) %,P <0.05].Before treatment,the FEV1,FEV1/FVC,PEF,the IL-17A,CXCL12 levels and TRPC1 level had no statistically significant differences between the two groups(P > 0.05).After treatment,the peripheral blood IL-17A [(24.18 ± 3.69) pg/mL],CXCL12 levels [(193.50 ± 2.90) pg/mL] and TRPC1 level in BALF [(7.69 ± 1.14)ng/L] in the observation group decreased significantly(P < 0.05),which in the control group also decreased significantly [IL-17A (34.25 ± 3.74) pg/mL,CXCL12 (205.37 ± 3.21) pg/mL,TRPC1 (14.25 ± 1.20)ng/L] (P < 0.05),which of the observation group were significantly lower than those of the control group(all P <0.05).The incidence rates of adverse effects of the two groups were t6.67%,12.82%,there was no statistically significant difference(P > 0.05).Conclusion Thymopentin combined with conventional therapy can effectively improve the elderly COPD patients'immunologic function and pulmonary ventilation function,decrease the inflammatory factor,relieve inflammatory reaction and it is safe and effective.

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