In vitro studies of H520 cell cycle and apoptosis by anlotinib combined with radiotherapy

Lu Guo, Luyao Zhang, Yan Guan, Yan Li, Chufeng Zhang, Qisen Guo, Lu Guo, Luyao Zhang, Yan Guan, Yan Li, Chufeng Zhang, Qisen Guo

Abstract

Background: To study in vitro the effects of anlotinib combined with radiotherapy on the lung cancer H520 cell cycle and apoptosis.

Methods: The log growth period H520 cells were divided into the control group, anlotinib group (A group), radiotherapy group (RT group) and combined group (A + RT group). Cell cycle and apoptosis were detected by flow cytometry and changes in H520 cell cycle and apoptosis were analyzed in each group.

Results: Anlotinib was determined to significantly inhibit cell growth in all groups, both alone, or in combination with radiotherapy. After receiving corresponding treatments, the proportions of G2/M-phase cells in the control group, A group, RT group and A + RT group were different, and statistically significant (F = 32.086, P < 0.001). The apoptotic cell statistics of H520 cells in the control group, A group, RT group and A + RT group were significantly different (F = 44.537, P < 0.01). The relative expression of CDK1 in each group of cells was 0.04 ± 0.02, 0.07 ± 0.12, 0.81 ± 0.11, and 0.56 ± 0.16, respectively. There were differences between the groups by analysis of variance which were statistically significant (F = 58.36, P < 0.0001). The relative expression of cycle B in each group of cells was 0.27 ± 0.05, 0.40 ± 0.16, 0.65 ± 0.14, and 0.57 ± 0.13, respectively. There were differences between the groups by analysis of variance which were statistically significant (F = 10.77, P = 0.0002).

Conclusions: Anlotinib had an inhibitory effect on lung cancer H520 cell proliferation. A higher rate of apoptosis and G2/M phase block was observed in the anlotinib-radiotherapy combined group. Anlotinib combined with radiotherapy was able to synergistically inhibit tumor cell growth.

Key points: Anrotinib combined with radiotherapy can synergistically inhibit tumor cell growth.

Keywords: Anlotinib; cell cycle/apoptosis; lung cancer; radiotherapy.

© 2021 The Authors. Thoracic Cancer published by China Lung Oncology Group and John Wiley & Sons Australia, Ltd.

Figures

Figure 1
Figure 1
(a) Inhibitory effect of anlotinib on H520, H226 and H2170 cells () Anlotinib, () Combined, () Anlotinib, () Combined, () Anlotinib, () Combined. (b) Inhibitory effect of anlotinib on H520, H226 and H2170 cells () H520, () H226, () H2170.
Figure 2
Figure 2
(a) Inhibitory effect of anlotinib on H520 cells () 24 hours, () 48 hours, () 72 hours. (b) Inhibitory effect of anlotinib on H520 cells.
Figure 3
Figure 3
(a) Flow cytometric results of four groups. (b) Comparison of G2/M proportion in four groups.
Figure 4
Figure 4
(a) Apoptosis of cell lines in four groups. (b) Apoptosis of cell lines in four groups.
Figure 5
Figure 5
(a) Expression and statistical analysis of CDK1 in each group of cells. (b) Expression and statistical analysis of CDK1 in each group of cells () Control, () RT, () A, () Combined. (c) Expression and statistical analysis of cycle B in each group of cells. (d) Expression and statistical analysis of cycle B in each group of cells () Control, () RT, () A, () Combined.
Figure 6
Figure 6
(a) Expression and statistical analysis of caspase‐3 in each group of cells. (b) Expression and statistical analysis of caspase‐3 in each group of cells () Combined, () RT, () A, () Control.

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