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Table 2 Measurement of doxorubicin-induced MDA-MB-468 cell apoptosis and growth inhibition

From: Hyaluronan-CD44 interaction promotes c-Jun signaling and miRNA21 expression leading to Bcl-2 expression and chemoresistance in breast cancer cells

Treatments

Apoptotic cells (Annexin V-positive cell/total cells × 100%)*

Doxorubicin-induced tumor cell growth inhibition IC50(μM)**

 

No doxorubicin

+ doxorubicin

 

(A) Effects of various signaling perturbation agents on doxorubicin--induced apoptosis and cell growth inhibition in MDA-MB-468 cells.

(a) Effects of Doxorubicin-induced apoptosis and cell growth inhibition in MDA-MB-468 cells following HA treatment:

No HA (Untreated cells)

4.1 ± 0.5

30.2 ± 3.1a

73.5 ± 5.1

+ HA

2.4 ± 0.3a

12.6 ± 2.3a

188.0 ± 10.5b

Non-immune rat IgG-treated cells (No HA)

3.8 ± 0.5a

29.2 ± 2.2a

70.3 ± 4.1b

Non-immune rat IgG-treated cells (+ HA)

2.2 ± 0.2a

10.5 ± 2.0a

176.0 ± 10.3b

Anti-CD44-treated cells (+ HA)

3.8 ± 0.2a

29.2 ± 2.2a

28.4 ± 3.6b

(b) Effects of Doxorubicin-induced apoptosis and cell growth inhibition in MDA-MB-468 cells treated with c-Jun siRNA plus HA:

Scrambled siRNA-treated cells (No HA)

5.2 ± 0.6

32.4 ± 3.6c

85.0 ± 2.4

Scrambled siRNA-treated cells (+ HA)

4.5 ± 0.5c

11.9 ± 3.1c

190.0 ± 6.00d

c-Jun siRNA-treated cells (No HA)

5.2 ± 0.6c

33.6 ± 2.8c

40.5 ± 0.30d

c-Jun siRNA-treated cells (+ HA)

4.5 ± 0.5c

31.9 ± 3.2c

39.2 ± 0.80d

(c) Effects of Doxorubicin-induced apoptosis and cell growth inhibition in MDA-MB-468 cells treated with anti-miR-21 inhibitor plus HA:

miRNA negative control-treated cells (No HA)

3.7 ± 0.4

31.6 ± 2.9e

62.8 ± 0.5

miRNA negative control-treated cells (+ HA)

2.5 ± 0.2e

12.3 ± 0.4e

196.0 ± 4.2f

Anti-miR-21 inhibitor-treated cells (No HA)

3.2 ± 0.6e

38 ± 2.5e

65.0 ± 3.0f

Anti-miR-21 inhibitor-treated cells (+ HA)

3.1 ± 0.4e

34 ± 2.1e

70.0 ± 0.2f

(B) Effects of JNK inhibitor (420116) on Doxorubicin-induced apoptosis and cell growth inhibition in MDA-MB-468 cells.

Vehicle control-treated cells (No HA)

4.3 ± 0.2

32 ± 2.4g

64.00 ± 6.2

Vehicle control-treated cells (+ HA)

2.6 ± 0.4g

14 ± 0.5g

180.0 ± 10.0h

JNK inhibitor (420116)-treated cells (No HA)

12.0 ± 0.5g

35 ± 2.6g

26.0 ± 3.5h

JNK inhibitor (420116)-treated cells (+ HA)

3.2 ± 0.3g

38 ± 3.3g

25.0 ± 0.2h

  1. *Cells were designated apoptotic when displaying Annexin V-positive staining. In each sample, at least 500 cells from five different fields were counted, with the percentage of Doxorubicin or JNK inhibitor (420116) (2 h treatment)-induced apoptotic cells calculated as Annexin V-positive cells/total number of cells. The values are presented as the means ± standard deviation. All assays consisted of at least six replicates and were performed on at least 4 different experiments.
  2. ** Tumor cell growth inhibition (IC50) is designated as “the μM concentration of chemotherapeutic drug (e.g., Doxorubicin-24 h treatment) that causes 50% inhibition of tumor cell growth” using MTT-based growth assay as described in the Materials and methods. IC50 values are presented as the means ± standard deviation. All assays consisted of at least six replicates and were performed on at least 4 different experiments.
  3. a, bStatistically significant (P < 0.005; analysis of variance; n = 4) as compared with control samples (No HA treatment).
  4. c, dStatistically significant (P < 0.005; analysis of variance; n = 4) as compared with control samples (scrambled siRNA-treated cells without HA addition).
  5. e, fStatistically significant (P < 0.005; analysis of variance; n = 4) as compared with control samples (miRNA negative control-treated cells without HA addition).
  6. g, hStatistically significant (P < 0.001; analysis of variance; n = 4) as compared with control samples (vehicle control-treated cells without HA addition).