Mechanism of miR-214 on the proliferation and apoptosis of oropharynx squamous cell carcinoma cell line by regulating Notch1 signal
Shaobin Liu, Zichen Liu, Changjuan Meng, Cuili Zhang
Abstract
Shaobin Liu, Zichen Liu, Changjuan Meng, Cuili Zhang
Abstract
Objective To study the mechanism of miR-214 on the proliferation, invasion and apoptosis of oropharyngeal squamous cell carcinoma (OSCC) cell line by regulating Notch 1 signal. Methods OSCC cell lines were purchased from ATCC cell bank and divided into three groups: inhibition group (IN group) , simulation group (SI group) and control group (CO group) . Notch 1 protein, mRNA expression, apoptosis, migration and proliferation were analyzed by Western blot, qRT-PCR, TUNEL, invasion experiment and CCK-8. One way ANOVA was used to compare differences among the three groups, LSD-t test was used for comparison between two groups. Result The expression of mir-214m RNA in the cells of IN group, SI group and CO group were (1.15 ±0.26) , (3.34±0.89) , (2.08±0.67) respectively, and there was significant difference between the SI group and IN group, SI group and CO group (t = 5.281, P = 0.002; t = 2.529, P = 0.045) ; The apoptosis rates of OSCC cells in the three groups were (0.78±0.10) ﹪, (0.32±0.06) ﹪, (0.56±0.12) ﹪, respectively. The difference between IN group and CO group, IN group and SI group were statistically significant (t = 3.149, P = 0.020; t = 8.823, P < 0.001) ; The proliferation rates of cancer cells in the three groups were: (0.65±0.05) ﹪, (1.26±0.06) ﹪, (1.89±0.04) ﹪, respectively. The difference between IN group and SI group, IN group and CO group were statistically significant (t = 11.056, P < 0.001; t = 27.394, P < 0.001) . The difference between CO group and SI group was statistically significant (t = 12.365, P < 0.001) ; the invasive rates of cancer cells in the three groups were: (0.13±0.02) ﹪, (0.89±0.13) ﹪, (0.48±0.11) ﹪, respectively. The difference between SI group and CO group, SI group and IN group were tatistically significant (t = 4.176, P = 0.006; t = 10.147, P < 0.001) ; The levels of Notch1 protein in the three groups were (2.38±1.34) , (0.94 ± 0.23) , (1.76 ± 0.67) , respectively. There was a significant difference between SI group and CO group (t = 2.588, P = 0.041; t = 2.368, P = 0.056) ; The expression of Notch1 mRNA in the three groups were (4.26 ± 1.06) , (1.45 ± 0.38) , (2.46 ± 0.87) , respectively. The difference was statistically significant (t = 2.935, P = 0.026; t = 5.583, P = 0.001) . Conclusion miR-214 levels are elevated and may be inversely related to the proliferation of OSCC cell lines. MiR-214 inhibits Notch1 signaling pathway expression, promotes growth and invasion of cancer cells, and inhibites apoptosis of cancer cells. Key words: miR-214; Notch1; Oropharyngeal squamous cell carcinoma
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Objective To study the mechanism of miR-214 on the proliferation, invasion and apoptosis of oropharyngeal squamous cell carcinoma (OSCC) cell line by regulating Notch 1 signal. Methods OSCC cell lines were purchased from ATCC cell bank and divided into three groups: inhibition group (IN group) , simulation group (SI group) and control group (CO group) . Notch 1 protein, mRNA expression, apoptosis, migration and proliferation were analyzed by Western blot, qRT-PCR, TUNEL, invasion experiment and CCK-8. One way ANOVA was used to compare differences among the three groups, LSD-t test was used for comparison between two groups. Result The expression of mir-214m RNA in the cells of IN group, SI group and CO group were (1.15 ±0.26) , (3.34±0.89) , (2.08±0.67) respectively, and there was significant difference between the SI group and IN group, SI group and CO group (t = 5.281, P = 0.002; t = 2.529, P = 0.045) ; The apoptosis rates of OSCC cells in the three groups were (0.78±0.10) ﹪, (0.32±0.06) ﹪, (0.56±0.12) ﹪, respectively. The difference between IN group and CO group, IN group and SI group were statistically significant (t = 3.149, P = 0.020; t = 8.823, P < 0.001) ; The proliferation rates of cancer cells in the three groups were: (0.65±0.05) ﹪, (1.26±0.06) ﹪, (1.89±0.04) ﹪, respectively. The difference between IN group and SI group, IN group and CO group were statistically significant (t = 11.056, P < 0.001; t = 27.394, P < 0.001) . The difference between CO group and SI group was statistically significant (t = 12.365, P < 0.001) ; the invasive rates of cancer cells in the three groups were: (0.13±0.02) ﹪, (0.89±0.13) ﹪, (0.48±0.11) ﹪, respectively. The difference between SI group and CO group, SI group and IN group were tatistically significant (t = 4.176, P = 0.006; t = 10.147, P < 0.001) ; The levels of Notch1 protein in the three groups were (2.38±1.34) , (0.94 ± 0.23) , (1.76 ± 0.67) , respectively. There was a significant difference between SI group and CO group (t = 2.588, P = 0.041; t = 2.368, P = 0.056) ; The expression of Notch1 mRNA in the three groups were (4.26 ± 1.06) , (1.45 ± 0.38) , (2.46 ± 0.87) , respectively. The difference was statistically significant (t = 2.935, P = 0.026; t = 5.583, P = 0.001) . Conclusion miR-214 levels are elevated and may be inversely related to the proliferation of OSCC cell lines. MiR-214 inhibits Notch1 signaling pathway expression, promotes growth and invasion of cancer cells, and inhibites apoptosis of cancer cells. Key words: miR-214; Notch1; Oropharyngeal squamous cell carcinoma
Key concepts: Apoptosis, Cell growth, Western blot, Cell culture, TUNEL assay, Biology, Group A, Molecular biology