Differential effects of sevoflurane on the growth and apoptosis of human cancer cell lines

[1] Robinson BJ, Uhrich TD, Ebert TJ. A review of recovery from sevoflurane anaesthesia:

comparisons with isoflurane and propofol including meta-analysis. Acta Anaesthesiol

Scand. 1999;43(2):185-90.

[2] Tavare AN, Perry NJS, Benzonana LL, Takata M, Ma D. Cancer recurrence after

surgery:

Direct

and

indirect

effects

of

anesthetic

agents.

Int

Cancer.

2012;130(6):1237-50.

[3] Brioni JD, Varughese S, Ahmed R, Bein B. A clinical review of inhalation anesthesia

with sevoflurane: from early research to emerging topics. J Anesth. 2017; 31( 5): 764-78.

[4] Ecimovic P, McHugh B, Murray D, Doran P, Buggy DJ. Effects of sevoflurane on breast

cancer cell function in vitro. Anticancer Res. 2013;33(10):4255-60.

[5] Jaura AI, Flood G, Gallagher HC, Buggy DJ. Differential effects of serum from patients

administered distinct anaesthetic techniques on apoptosis in breast cancer cells in vitro:

a pilot study. Br J Anaesth. 2014;113 Suppl 1:i63-7.

[6] Nishiwada T, Kawaraguchi Y, Uemura K, Sugimoto H, Kawaguchi M. Effect of

sevoflurane on human hepatocellular carcinoma HepG2 cells under conditions of high

glucose and insulin. J Anesth. 2015;29(5):805-8.

[7] Shi, Q. Y., S. J. Zhang, L. Liu, Q. S. Chen, L. N. Yu, F. J. Zhang and M. Yan.

Sevoflurane promotes the expansion of glioma stem cells through activation of

hypoxia-inducible factors in vitro. Br J Anaesth. 2015;114(5):825-30.

[8] Benzonana, L. L., N. J. Perry, H. R. Watts, B. Yang, I. A. Perry, C. Coombes, M. Takata

and D. Ma. Isoflurane, a commonly used volatile anesthetic, enhances renal cancer

growth and malignant potential via the hypoxia-inducible factor cellular signaling

pathway in vitro. Anesthesiology. 2013;119(3):593-605.

[9] Kawaraguchi, Y., Y. T. Horikawa, A. N. Murphy, F. Murray, A. Miyanohara, S. S. Ali, B.

P. Head, P. M. Patel, D. M. Roth and H. H. Patel. Volatile anesthetics protect cancer

cells against tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis

via caveolins. Anesthesiology. 2011;115(3):499-508.

[10] Niwa H, Rowbotham DJ, Lambert DG, Buggy DJ. Can anesthetic techniques or drugs

affect cancer recurrence in patients undergoing cancer surgery? J Anesth.

2013;27(5):731-41.

[11] Kvolik S, Glavas-Obrovac L, Bares V, Karner I. Effects of inhalation anesthetics

halothane,

sevoflurane,

and

isoflurane

on

human

cell

lines.

Life

Sci.

2005;77(19):2369-83.

[12] Kvolik S, Dobrosevic B, Marczi S, Prlic L, Glavas-Obrovac L. Different apoptosis ratios

21

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

and gene expressions in two human cell lines after sevoflurane anaesthesia. Acta

Anaesthesiol Scand. 2009;53(9):1192-9.

[13] Muller-Edenborn, B., B. Roth-Z'graggen, K. Bartnicka, A. Borgeat, A. Hoos, L. Borsig

and B. Beck-Schimmer. Volatile anesthetics reduce invasion of colorectal cancer cells

through

down-regulation

of

matrix

metalloproteinase-9.

Anesthesiology.

2012;117(2):293-301.

[14] Liang H, Gu M, Yang C, Wang H, Wen X, Zhou Q. Sevoflurane inhibits invasion and

migration of lung cancer cells by inactivating the p38 MAPK signaling pathway. J

Anesth. 2012;26(3):381-92.

[15] Liu S, Fang F, Song R, Gao X, Jiang M, Cang J. Sevoflurane affects neurogenesis

through cell cycle arrest via inhibiting wnt/beta-catenin signaling pathway in mouse

neural stem cells. Life Sci. 2018;209:34-42.

[16] Loop, T., D. Dovi-Akue, M. Frick, M. Roesslein, L. Egger, M. Humar, A. Hoetzel, R.

Schmidt, C. Borner, H. L. Pahl, K. K. Geiger and B. H. Pannen. Volatile anesthetics

induce caspase-dependent, mitochondria-mediated apoptosis in human T lymphocytes

in vitro. Anesthesiology. 2005;102(6):1147-57.

[17] Dong, Y., G. Zhang, B. Zhang, R. D. Moir, W. Xia, E. R. Marcantonio, D. J. Culley, G.

Crosby, R. E. Tanzi and Z. Xie. The common inhalational anesthetic sevoflurane

induces

apoptosis

and

increases

beta-amyloid

protein

levels.

Arch

Neurol.

2009;66(5):620-31.

[18] Topouzova-Hristova T, Daza P, Garcia-Herdugo G, Stephanova E. Volatile anaesthetic

halothane causes DNA damage in A549 lung cells. Toxicol In Vitro. 2006;20(5):585-93.

[19] Yang, Zeyong, Jingjing Lv, Xingxing Li, Qiong Meng, Qiling Yang, Wei Ma, Yuanhai Li

and Zun Ji Ke. Sevoflurane decreases self-renewal capacity and causes c-Jun

N-terminal kinase–mediated damage of rat fetal neural stem cells. Sci Rep.

2017;7:46304.

[20] Kuilman, T., C. Michaloglou, L. C. Vredeveld, S. Douma, R. van Doorn, C. J. Desmet, L.

A. Aarden, W. J. Mooi and D. S. Peeper. Oncogene-induced senescence relayed by an

interleukin-dependent inflammatory network. Cell. 2008;133(6):1019-31.

[21] Dhanasekaran

DN,

2008;27(48):6245-51.

Reddy

EP.

JNK

signaling

in

apoptosis.

Oncogene.

22

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

Figure legends

Figure 1:

Figure 1. Differential effects of 1% sevoflurane on the growth of human cancer cell

lines.

(a) Experimental design. Cells were exposed to 1% (v/v) sevoflurane in air for the

duration indicated by grey bars, incubated for 48 h, and then enumerated.

(b) Proliferation indices at 48 h after cessation of exposure in nine human cell lines. The

data for each exposure time are shown relative to the non-exposure control (mean ± SD;

n = 6). * P < 0.05; ** P < 0.01.

Figure 2:

Figure 2. Enhanced Matrigel invasion in cancer cell lines growth-promoted but not

growth-suppressed by 1% sevoflurane.

(a) Experimental design of Boyden chamber-based invasion assays. Cells were

incubated with or without 1% (v/v) sevoflurane for 4 h, seeded into Transwell inserts

coated with Matrigel, and allowed to migrate into Matrigel for 24 h.

(b) Representative images of crystal violet-stained cells that have migrated through

Matrigel. Original magnification, 40×; scale bar, 50 μm.

23

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

(c) Summary of the results. For each sample, migrating cells were enumerated in at least

five visual fields under microscopy, and a mean cell number per visual field was

determined. Data are expressed as mean ± SD values in biological triplicates for each

experimental condition. ** P < 0.01.

Figure 3:

Figure 3. Induction of early apoptosis in cancer cell lines growth-suppressed but

not growth-promoted by 1% sevoflurane.

(a) Experimental design. Cells were exposed to 1% (v/v) sevoflurane for the duration

indicated by grey bars, incubated for 24 h, stained with Annexin V, and then analyzed

via flow cytometry.

(b) Quantification of FITC-conjugated Annexin V fluorescence. The data for each

exposure time are shown relative to the non-exposure control (mean ± SD; n = 4). * P <

0.05; ** P < 0.01.

(c) Representative histograms of four cell lines.

Figure 4:

Figure 4. Enhanced expression of cleaved caspase-3 in cancer cell lines

24

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

growth-suppressed but not growth-promoted by 1% sevoflurane.

(a) Experimental design. Cells were exposed to 1% (v/v) sevoflurane for the duration

indicated by grey bars, incubated for 24 h, and harvested. Cell lysates were examined

using immunoblotting against caspase-3 and β-actin. β-Actin was used as a loading

control.

(b) Images of immunoblotting results. Bands for intact caspase-3, cleaved caspase-3,

and β-actin are indicated on the left. Half the cell samples were incubated in the

presence of pan-caspase inhibitor Z-VAD-FMK during 24 h incubation so that the

depletion of cleaved caspase-3 could help identify the corresponding signal.

Figure 5:

Figure 5. Induction of cell death in cancer cell lines growth-suppressed but not

growth-promoted by 1% sevoflurane.

(a) Experimental design. Cells were exposed to 1% (v/v) sevoflurane for the duration

indicated by grey bars, incubated for 24 h, stained with 7-AAD, and analyzed via flow

cytometry.

(b) Quantification of 7-AAD fluorescence. The data for each exposure time are shown

relative to the non-exposure control (mean ± SD; n = 4). ** P < 0.01.

25

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

(c) Representative histograms of four cell lines.

Figure

Figure

Figure 2

NCI-H1299

Control

Sevo 4 h

A549

Control

Sevo 4 h

MDA-MB-231

Control

Sevo 4 h

MCF7

Control

Sevo 4 h

Number of migrating cells

NCI-H1299

A549

MDA-MB-231

MCF7

Figure 3

Relative intensity (Annexin V)

Exposure time (1% v/v sevoflurane)

Intensity (Annexin V)

Exposure time (1% v/v sevoflurane)

Figure 4

Figure 5

Relative intensity (7-AAD)

Exposure time (1% v/v sevoflurane)

Intensity (7-AAD)

Exposure time (1% v/v sevoflurane)

...