World J Oncol

World Journal of Oncology, ISSN 1920-4531 print, 1920-454X online, Open Access

Article copyright, the authors; Journal compilation copyright, World J Oncol and Elmer Press Inc

Journal website https://www.wjon.org

Original Article

Volume 15, Number 1, February 2024, pages 38-44

Viability Profiles of Normal and Cancer Bladder Cells With Metformin, Nitrate and Adenosine Monophosphate-Activated Protein Kinase Inhibitor

Figure 1. Correlation between concentrations of NaNO₃ and cell viabilities of normal bladder cells (PdECs) after 48 h.

Figure 2. Comparison of changes in cell viability profiles of normal bladder cells (PdECs) with 300 µM NaNO₃ and 1.0 × 10³ µM metformin when treated alone or combined. Both 300 µM NaNO₃ and 1.0 × 10³ µM metformin were added to PdEC at the same time.

**Table 1.** Viability Profiles (%) of Normal Bladder Cells (PdECs) With Different Concentrations of NaNO₃ and Metformin After 48 h
Concentration, µM	NaNO₃	Metformin
Each value is the average values of three runs.
Control	82.3	66.4
30	75.7
60	69.6
90	63.4
200	55.4
300	33.5
500	0.0
1.0 × 10³		65.4
5.0 × 10³		63.1
1.0 × 10⁴		63.6

Table 1. Viability Profiles (%) of Normal Bladder Cells (PdECs) With Different Concentrations of NaNO₃ and Metformin After 48 h

Concentration, µM

NaNO₃

Metformin

Each value is the average values of three runs.

Control

82.3

66.4

75.7

69.6

63.4

200

55.4

300

33.5

500

0.0

1.0 × 10³

65.4

5.0 × 10³

63.1

1.0 × 10⁴

63.6

**Table 2.** Viability Profiles (%) of Normal Bladder Cells (PdECs) With 300 µM NaNO₃ and 1.0 × 10³ µM Metformin (Met) When Treated Alone or Combined
Concentration, µM	24 h	48 h
Each value is the average value of three runs.
Control	93.3	93.3	93.3	86.0	86.0	86.0
300 µM NaNO₃	66.7			42.7
1.0 × 10³ µM metformin		92.7			84.3
Both			77.0			69.3
Δ in cell viability	26.6	0.6	16.3	43.3	1.7	16.7

Table 2. Viability Profiles (%) of Normal Bladder Cells (PdECs) With 300 µM NaNO₃ and 1.0 × 10³ µM Metformin (Met) When Treated Alone or Combined

Concentration, µM

24 h

48 h

NaNO₃, %

Met, %

Both, %

NaNO₃, %

Met, %

Both, %

Each value is the average value of three runs.

Control

93.3

86.0

300 µM NaNO₃

66.7

42.7

1.0 × 10³ µM metformin

92.7

84.3

Both

77.0

69.3

Δ in cell viability

26.6

0.6

16.3

43.3

1.7

16.7

**Table 3.** Viability Profiles (%) of Normal Bladder Cells (PdECs) With Sequential Treatments of NaNO₃, Metformin, and/or AMPK Inhibitor (Compound C) After 72 h
Addition order	Cell viability, %	Δ in cell viability, %^b
Each value is the average value of three runs. ^aIncubation time. ^bEach value is calculated as control minus the experiment.
(a) Control	92.0
(b) 300 µM NaNO₃	23.3	68.7
(c) 1.0 × 10³ µM metformin	91.7	0.3
(d) 1.0 × 10³ µM compound C	88.7	3.3
(e) 300 µM NaNO₃ (24 h)^a, 1.0 × 10³ µM metformin (48 h)^a	84.3	7.7
(f) 1.0 × 10³ µM metformin (24 h)^a, 300 µM NaNO₃ (48 h)^a	84.0	8.0
(g) 1.0 × 10³ compound C (6 h pre-treatment)^a, 300 µM NaNO₃ (24 h)^a, 1.0 × 10³ µM metformin (48 h)^a	55.3	36.7
(h) 1.0 × 10³ compound C (6 h pre-treatment)^a, 1.0 × 10³ µM metformin (24 h)^a, 300 µM NaNO₃ (48 h)^a	56.0	36.0

Table 3. Viability Profiles (%) of Normal Bladder Cells (PdECs) With Sequential Treatments of NaNO₃, Metformin, and/or AMPK Inhibitor (Compound C) After 72 h

Addition order

Cell viability, %

Δ in cell viability, %^b

Each value is the average value of three runs. ^aIncubation time. ^bEach value is calculated as control minus the experiment.

(a) Control

92.0

(b) 300 µM NaNO₃

23.3

68.7

91.7

0.3

(d) 1.0 × 10³ µM compound C

88.7

3.3

(e) 300 µM NaNO₃ (24 h)^a, 1.0 × 10³ µM metformin (48 h)^a

84.3

7.7

(f) 1.0 × 10³ µM metformin (24 h)^a, 300 µM NaNO₃ (48 h)^a

84.0

8.0

(g) 1.0 × 10³ compound C (6 h pre-treatment)^a, 300 µM NaNO₃ (24 h)^a, 1.0 × 10³ µM metformin (48 h)^a

55.3

36.7

(h) 1.0 × 10³ compound C (6 h pre-treatment)^a, 1.0 × 10³ µM metformin (24 h)^a, 300 µM NaNO₃ (48 h)^a

56.0

36.0

**Table 4.** Viability Profiles (%) of Cancer Bladder Cells (5367) With Different Concentrations of Metformin After 48 h
Concentration, µM	Cell viability, %	Δ in cell viability, %
Each value is the average value of three runs.
Control	91.3
1.0 × 10³	81.3	10.0
5.0 × 10³	74.9	16.4
1.0 × 10⁴	42.0	49.3

Table 4. Viability Profiles (%) of Cancer Bladder Cells (5367) With Different Concentrations of Metformin After 48 h

Concentration, µM

Cell viability, %

Δ in cell viability, %

Each value is the average value of three runs.

Control

91.3

1.0 × 10³

81.3

10.0

5.0 × 10³

74.9

16.4

1.0 × 10⁴

42.0

49.3