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Objective@#To elucidate the mechanism of melatonin combined with cisplatin in promoting cell apoptosis of rat pancreatic cancer AR42J cells.@*Methods@#Rat pancreatic cancer AR42J cells were divided into control group, 1 mmol/L cisplatin treated group (cisplatin group), 1 mmol/L melatonin treated group (melatonin group), 1 mmol/L cisplatin combined with 1 mmol/L melatonin treated group (combined group), 1 μmol/L cisplatin combined with melatonin treated group after 1 μmol/L PBN pretreatment for an hour (PBN+ combined group) and 1 μmol/L cisplatin combined with melatonin treated group after PBN solvent pretreatment for an hour (solvent+ combined group). MTT and annexin V-FITC/PI were used to detect the cell proliferation rate and cell apoptosis rate, respectively. The protein expression of caspase-3 was detected by Western blot. DCFH-DA was used to detect the level of ROS. ROS level and caspase-3 expression in AR42J cells pretreated with ROS antagonist PBN for 24 hours were detected.@*Results@#The cell proliferation rate of control group, cisplatin group, melatonin group and combination group after 24-hour culture was (96.29±3.49)%, (81.38±6.01)%, (80.72±3.68)% and (42.26±6.35)%, respectively. The cell apoptosis rate was (16.42±4.15)%, (56.47±9.06)%, (52.94±6.57)% and (87.36±6.48)%, respectively. The percentage of ROS positive cells was (1.33±1.53)%, (46.67±7.64)%, (45.67±5.13)% and (83.33±7.64)%, respectively. The expression of cspase-3 was 100%, (150.64±7.70)%, (147.00±7.27)% and (190.04±5.07)%, respectively. The cell proliferation rate of cisplatin group and melatonin group was significantly lower than that of the control group. The apoptotic rate, the proportion of ROS positive cells and the expression of caspase-3 were significantly higher than those in control group. The changes in the combined group were more obvious than those in the single drug treatment group, and the differences were all statistically significant (all Pvalues <0.05). The percentage of ROS positive AR42J cells [(45.24±4.64)% vs (84.90±6.13)%)] induced by melatonin combined with cisplatin could be significantly reversed by PBN pretreatment for 1 hour, and the expression of caspase-3 in AR42J cells could be down regulated (124.92±9.72 vs 184.69±7.76) with statistically significant difference (all P values<0.05).@*Conclusions@#Melatonin combined with cisplatin may promote cell apoptosis of pancreatic cancer AR42J cells through ROS/caspase-3 pathway.
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Objective To study the expression and significance of Nrf2 and caspase-3 in non-small cell lung cancer (NSCLC).Methods The specimens from 82 cases of NSCLC in our hospital from September 2015 to December 2017 were selected for observation.All patients were surgically resected to obtain NSCLC specimens.The expression of Nrf2 and caspase-3 in NSCLC was detected by immunohistochemistry and its clinical significance was analyzed.Results The positive expression rate of Nrf2 in NSCLC (63.4％) was significantly higher than that in normal tissue (30.0％),with statistically significant difference (P ＜ 0.05).The analysis of clinical characteristics showed that the expression of Nrf2 in NSCLC was related to tumor diameter,tumor node metastasis (TNM) stage,lymph node metastasis and distant metastasis (P ＜ 0.05).The expression of Nrf2 in NSCLC was not significantly associated with gender,age,smoking history,case classification,and histological differentiation (P ＞ 0.05).The positive rate of caspase-3 in NSCLC (40.2％) and the positive rate of caspase-3 in normal tissues (40.0％) were not statistically different (P ＞ 0.05).Analysis of clinical characteristics showed that the expression of caspase-3 in NSCLC was related to TNM stage,lymph node metastasis,distant metastasis,and the difference was statistically significant (P ＜ 0.05);the expression of Nrf2 in NSCLC was not significantly associated with gender,age,smoking history,case classification,tissue differentiation,and tumor diameter (P ＞ 0.05).Conclusions The abnormal expression of Nrf2 and caspase-3 is correlated with the progression of NSCLC and the malignant biological behavior.Active detection of Nrf2 and caspase-3 levels can clinically determine the clinical stage,lymph node metastasis and distant metastasis of NSCLC,providing information to facilitate clinical development of relevant interventions.
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Objective To elucidate the mechanism of melatonin combined with cisplatin in promoting cell apoptosis of rat pancreatic cancer AR42J cells. Methods Rat pancreatic cancer AR42J cells were divided into control group, 1 mmol/L cisplatin treated group (cisplatin group), 1 mmol/L melatonin treated group ( melatonin group) , 1 mmol/L cisplatin combined with 1 mmol/L melatonin treated group ( combined group) , 1 μmol/L cisplatin combined with melatonin treated group after 1 μmol/L PBN pretreatment for an hour ( PBN+combined group ) and 1 μmol/L cisplatin combined with melatonin treated group after PBN solvent pretreatment for an hour ( solvent+combined group ) . MTT and annexin V-FITC/PI were used to detect the cell proliferation rate and cell apoptosis rate, respectively. The protein expression of caspase-3 was detected by Western blot. DCFH-DA was used to detect the level of ROS. ROS level and caspase-3 expression in AR42J cells pretreated with ROS antagonist PBN for 24 hours were detected. Results The cell proliferation rate of control group, cisplatin group, melatonin group and combination group after 24-hour culture was (96. 29 ± 3. 49)％, (81. 38 ± 6. 01)％, (80. 72 ± 3. 68)％ and (42. 26 ± 6. 35)％, respectively. The cell apoptosis rate was (16. 42 ± 4. 15)％, (56. 47 ± 9. 06)％, (52. 94 ± 6. 57)％ and (87. 36 ± 6. 48)％, respectively. The percentage of ROS positive cells was (1. 33 ± 1. 53)％, (46. 67 ± 7. 64)％, (45. 67 ± 5. 13)％ and (83. 33 ± 7. 64)％, respectively. The expression of cspase-3 was 100％, (150. 64 ± 7. 70)％, (147. 00 ± 7. 27)％ and (190. 04 ± 5. 07)％, respectively. The cell proliferation rate of cisplatin group and melatonin group was significantly lower than that of the control group. The apoptotic rate, the proportion of ROS positive cells and the expression of caspase-3 were significantly higher than those in control group. The changes in the combined group were more obvious than those in the single drug treatment group, and the differences were all statistically significant (all Pvalues <0.05). The percentage of ROS positive AR42J cells [(45.24 ± 4. 64)％ vs (84. 90 ± 6. 13)％)] induced by melatonin combined with cisplatin could be significantly reversed by PBN pretreatment for 1 hour, and the expression of caspase-3 in AR42J cells could be down regulated (124. 92 ± 9. 72 vs 184. 69 ± 7. 76) with statistically significant difference (all P values<0. 05). Conclusions Melatonin combined with cisplatin may promote cell apoptosis of pancreatic cancer AR42J cells through ROS/caspase-3 pathway.
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Objective To study the biological effect of different dose of satereotactic body radiation therapy (SBRT) on hepatoma Hep G2 Cells.Methods Hep G2 hepatumor cells vitality of 0,2,4,6,8,and 10 Gy doses of SBRT were determined by methyl thiazolyl tetrazolium (MTT) method.The cell colony formation rate and the cell survival fraction were detected.The cell apoptosis was detected by flow cytometry.The expressions of cyclin D1,Bax,Bcl-2,and caspase-3 protein were detected by Western blot.Results Hep G2 cells vitality was obvious inhibitory effect in different doses of SBRT,increasing with the SBRT dose,the cell vitality inhibition [2 Gy group (4.29 ± 0.63) ％,4 Gy group (19.37 ± 4.54) ％,6 Gy group (41.35 ±4.51)％,8 Gy group (86.34 ±5.68)％,10 Gy group (97.24 ±5.41)％] was increased,compared to 0 Gy group [(2.94 ± 0.52) ％],the difference was statistically significant (all P ＜ 0.01 or P ＜0.05).The cell colony formation rate and survival score in different doses of SBRT were statistically lower than the control group,increasing with the SBRT dose,the cell colony formation rate [4 Gy group (83.15 ± 4.71),6 Gy group (65.23 ± 5.47),8 Gy group (41.28 ± 5.36),10 Gy group (8.14 ± 5.22)] and survival score [2 Gy group (95.49 ±3.21),4 Gy group (84.23 ±3.45),6 Gy group (66.08 ± 3.71),8 Gy group (41.82 ± 3.82),10 Gy group (8.25 ± 3.56)] were decreased,compared to 0 Gy group [(98.72 ± 1.23),(100.00 ±0.00)],the differences were statistically significant (all P ＜0.01 or P ＜ 0.05).The cell apoptosis in different doses of SBRT was increased,increasing with the SBRT dose,the Hep G2 cells apoptosis [4 Gy group (13.67 ±4.62),6 Gy group (28.71 ±5.13),8 Gy group (54.16 ±5.24),10 Gy group (76.31 ± 5.17)] were significantly increased,compared to 0 Gy group (1.35 ±0.46),the difference was statistically significant (all P ＜ 0.01).The expressions of cyclinD1,Bax,Bcl-2,and caspase-3 protein in different doses of SBRT were decreased,increasing with the SBRT dose,the cyclin D1 [6 Gy group (1.94 ± 0.57),8 Gy group (1.38 ± 0.41),10 Gy group (0.52 ± 0.34)],Bax[6 Gy group (2.67 ±0.59),8 Gy group (1.73 ±0.45),10 Gy group (0.63 ±0.37)],Bcl-2 [4 Gy group (1.78 ±0.41),6 Gy group (1.03 ±0.35),8 Gy group (0.76 ±0.28),10 Gy group (0.38 ± 0.21)],Caspase-3 [6 Gy group (0.76 ± 0.31),8 Gy group (0.42 ± 0.24),and 10 Gy group (0.24 ± 0.21)] were obviously decreased,compared to 0 Gy group [(3.41 ± 0.75),(4.36 ± 0.87),(2.57 ± 0.46),(1.67 ± 0.55)],the difference was statistically significant (all P ＜ 0.05).Conclusions Different dose of SBRT can reduce Hep G2 cell vitality,enhance radiation sensitivity,have a dose dependent,maybe related to promote cell apoptosis and inhibit the proliferation of protein expression.
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Objective To study the biological effects of dose radiotherapy for primary liver cancer patients with stereotactic body portion.Methods 80 patients with primary liver cancer were selected.According to a random number table method, all patients were divided into observation group and control group,40 patients in each group.All patients were treated with stereotactic body radiotherapy treatment, the observation group was treated with high dose, while the control group received low-dose.The radiotherapy effects of different doses were analyzed and compared.Results After treatment, the effective rate of the observation group was 77.5%,which of the control group was 55.0%, ,the difference between the two groups was statistically significant (x2=4.528,P=0.033).After treatment for 6 months,1 year and 2 years, the survival rates of the observation group were 77.5%,75.0%,72.5%, which were significantly higher than 55.5%,52.5%,50.0% of the control group (P<0.05).The incidence rate of adverse reactions of the observation group was 42.5%, which was significantly lower than 72.5% of the control group (x2=7.365,P=0.006).Conclusion Stereotactic body radiotherapy treatment with different therapeutic dose has different treatment efficacy, high-dose SBRT treatment has significantly higher efficacy, survival rate and lower adverse reactions, which is worthy of clinical application.