ABSTRACT
Objective:To explore the inhibitory effects of gefitinib (epidermal growth factor receptor inhibitor) com-bined with celecoxib (cyclooxygenase-2 inhibitor) against human lung cancer A549 cells and the possible mechanism. Methods: A549 cells were cultured in RPMI 1640 medium and were divided into 4 groups: normal control group, 5 μmoL/L gefitinib group, 25 μmol/L celecoxib group, and 5 μ mol/L gefitinib + 25 μmol/L celecoxib group. The morpho-logical changes of A549 cells were observed under inverted microscope 48 h after treatment ; the effects of drugs on growth of A549 Cells were detected by MTT assay; the apoptosis and cell cycles of A549 cells were measured by Annexin V/PI and Hoechst 33258 staining, respectively; and the expression of EGFR protein, COX-2 protein, and EGFR mRNA were determined by immunofluorescenee and real-time PCR. Results: Compared with gefitinib and celecoxib groups, many granules and vacuoles were observed in the gefitinib and celecoxib combination group, and cells became round and there was defluxion. Both gefitinib and celecoxib inhibited the growth of A549 cells in a time- and dose-dependent manner. Af-ter treatment for 48 h, the inhibitory rate was (58.2±4.6) % in the combination group, which was significantly higher than those of the other two groups. Apoptosis rate in the combination group was also significantly higher than those in the other two groups (33.9% vs 6.0%, 8.8%), and the cell proportion in S phase significantly decreased and in G_0/G_1 phases significantly increased(P <0.01). EGFR protein, COX-2 protein, and EGFR mRNA expression in A549 cells was significantly decreased in the combination treatment group compared with those in the other two groups (P < 0.05). Conclusion : Gefitinib and celecoxib can synergistically inhibit the growth of A549 cells, possibly through promoting apop-tosis, G_0/G_1 arrest, and down-regulating activated EGFR and COX-2 expression.
ABSTRACT
Objective:To explore the inhibitory effects of gefitinib(epidermal growth factor receptor inhibitor) combined with celecoxib(cyclooxygenase-2 inhibitor) against human lung cancer A549 cells and the possible mechanism.Methods: A549 cells were cultured in RPMI 1640 medium and were divided into 4 groups: normal control group,5 ?mol/L gefitinib group,25 ?mol/L celecoxib group,and 5 ?mol/L gefitinib+25 ?mol/L celecoxib group.The morphological changes of A549 cells were observed under inverted microscope 48 h after treatment;the effects of drugs on growth of A549 Cells were detected by MTT assay;the apoptosis and cell cycles of A549 cells were measured by Annexin V/PI and Hoechst 33258 staining,respectively;and the expression of EGFR protein,COX-2 protein,and EGFR mRNA were determined by immunofluorescence and real-time PCR.Results: Compared with gefitinib and celecoxib groups,many granules and vacuoles were observed in the gefitinib and celecoxib combination group,and cells became round and there was defluxion.Both gefitinib and celecoxib inhibited the growth of A549 cells in a time-and dose-dependent manner.After treatment for 48 h,the inhibitory rate was(58.2?4.6)% in the combination group,which was significantly higher than those of the other two groups.Apoptosis rate in the combination group was also significantly higher than those in the other two groups(33.9% vs 6.0%,8.8%),and the cell proportion in S phase significantly decreased and in G0/G1 phases significantly increased(P
ABSTRACT
BACKGROUND:Idiopathic interstitial pneumonia is of poor response to treatment. Glucocorticoids are the first medicine for the treatment, however there is only 30% of the patients who are responded. Traditional Chinese drugs (TCD) have been researched hot point for prevention and treatment of pulmonary fibrosis. Many TCD have been used clinically, and with a certain therapeutic effect. Transforming growth factor-β1 and tumor necrosis factor-α are the considerable cytokines to cause pulmonary fibrosis, inhibition of their expression, therefore, may be effective to pulmonary fibrosis.OBJECTIVE: To investigate the interventional effect of qidan granule on pulmonary fibrosis in rats induced by bleomycin A5 and the influence on the expressions of transforming growth factor-β1 and tumor necrosis factorα, and also to compare with those of hydrocortisone.DESIGN:A randomized and interval grouping design.SETTING:Department of Respiratory Medicine, Shandong Provincial Hospital, Shandong University.MATERIALS:The experiment was conducted from May 2003 to March 2004 at Pathological Laboratory of Shandong Provincial Academy of Medical Science. Totally 105 SD male rats, were at random divided into 4groups: normal control group (n= 15 ), model group, qidan group and hydrocortisone group, with 30 rats in each group. Each group was subdivided as7-day, 14-day and 28-day group, with 5 rats in each normal group, and 10in each other groups.METHODS: [1] Model establishment: A perfusion was intrabronchially performed, of 0.25 mL normal saline for rats in normal control group, and of bleomycin A5 0.25 mL ( 5 mg/kg,4 g/L) for rats in other 3 groups, to set up the models of pulmonary fibrosis. [2]Administration: Next day to the beginning of modeling qidan granule (consisting of Radix Astragali seu Hedysari, Radix Salviae Miltiorrhizae, Rhizoma Ligustici Chuanxiong and so on, 3 125 mg/kg) was intragastrically given per day for rats in qidan group, hydrocortisone (25 mg/kg) was intraperitoneally given per day for rats in hydrocortisone group, and normal saline (2 mL/rat) was intragastrically given per day for rats in normal and model groups.[3] Observation indexes: The rats in each group were on the day 7, day 14 and day 28 put to death under the anesthesia, then the lung tissue was taken, stained with hematoxyline-eosin stain for pathological observation of lung tissue. The expressions of transforming growth factor-β1 and tumor necrosis factor-α were detected by immunohistochemistry.MAIN OUTCOME MEASURES:Pathological observation of lung tissue,and the expressions of transforming growth factor-β1 and tumor necrosis factor-α at different time points of rats in each group.RESULTS:Totally 100 rats entered the final result analysis.[1]Pathological observation of lung tissue: In the normal group the structure was normal, in the model group there were alveolitis on the day 7, deterioration of alveolitis on the day 14, and extensive fibrosis on the day 28; the degrees of alveolitis and fibrosis in the qidan group were slighter than those in the model group, and there was normal structure of alveoli; and in the hydrocortisone group the alveolitis on the day 7 and 14 was slighter than that in the model group, but there was no significant difference of fibrosis compared with the model group.[2] Expression of transforming growth factor-β1:In the model group the expression was highest on the day 28 and obviously higher than that in the normal group (3.6±0.4,1.2±0.4,P ＜ 0.01 ); the expression in the qidan group and hydrocortisone group was obviously lower than that in the model group(1.7±0.5,2.5±0.4,P ＜ 0.01), and the expression in the qidan group was lower than that in the hydrocortisone group (P＜ 0.01 ). [3]Expression of tumor necrosis factor-α: In the model group at different time points the expression was continuously increased, the expression in the qidan group and hydrocortisone group was obviously lower than that in the model group(P ＜ 0.05 or P ＜ 0.01), and the expression in the qidan group was lower than that in the hydrocortisone group ( P ＜ 0.01).CONCLUSION: Qidan granule can obviously reduce the extent of pulmonary fibrosis in rats induced by bleomycin A5, lower the expressions of transforming growth factor-β1 and tumor necrosis factor-α, and the effect was better than that of hydrocortisone.