Heteronemin, a Spongean Sesterterpene, Induces Cell Apoptosis and Autophagy in Human Renal Carcinoma Cells

Figure 7

Inhibition of autophagy enhanced the anticancer effect of heteronemin in A498 cells. A498 cells were pretreated with autophagy inhibitor, chloroquine, for 30 min, then 3 μM heteronemin was added for 24 h, and (a) the cell viability was determined using MTT assay. A498 cells were transfected with Atg5 siRNA or negative control and (b) the cell viability was determined using MTT assay and (c) the expression of apoptosis-related proteins (PARP and procaspase-3) and autophagy-related proteins (LC3 and Atg5) was evaluated for 24 h by western blotting. A498 cells were pretreated with JNK inhibitor, SP600125, for 30 min, then 3 μM heteronemin was added for 24 h, and (d) the cell viability was determined using MTT assay and (e) the expression of apoptosis-related proteins (PARP and procaspase-3) and LC3 was evaluated for 24 h by western blotting. H, CQ, and SP are indicated as heteronemin 3 μM, chloroquine 50 μM, and SP600125 20 μM, respectively. compared with the control group. compared with the heteronemin-treated group. CTL is indicated as control. DMSO was used as the vehicle control (CTL).

(a)

(b)

(c)

(d)

(e)