## 2 Results
### Abietic Acid Modulates Key- Genes Regulating Multiple Controlling Pathways
This may suggest the potential role of both pine extract and abiotic acid in apoptosis through angiogenesis and carcinogenesis. It was noted that kinases play important roles as molecular therapeutic targets in modern anticancer therapy. Pine extract and abiotic acid were shown to downregulate the level of _PKC-\(\alpha\)_ after 4 h, with superior significant reduction for abiotic acid-treated cells. This effect lasted at a minimum level after 24 h. Another key kinase in MCF-7 resistance and metastasis is _PRKAA1_ (Protein Kinase AMP-Activated Catalytic Subunit Alpha 1); both pine extract and abiotic acid were able to overexpress _PRKAA1_, which activated AMPK after 4 h and increased in a time-dependent manner after 24 h. Finally, pine extract was shown to slightly induce the overexpression of _CDK-4_ after 8 h, but downregulation occurred after 24 h, while abiotic acid reduced _CDK-4_ gene levels after 24 h.  
Abietic Acid Induces Increased Protein Level of Cytochrome-C Confirmed by Immunocytochemistry (ICC) Analysis  
The apoptotic effect was morphologically confirmed, as shown in Figure 2A. Cells treated with IC\({}_{50}\) of pine extract and abiotic acid showed similar characteristic features of \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline
**Genes** & **Fine Total Extract-RQ, Fold-Change (Mean \(\pm\) SEM)** & \multicolumn{3}{c}{**Abietic Acid-RQ, Fold-Change (Mean \(\pm\) SEM)**} \\ \hline  & **4 h** & **8 h** & **24 h** & **4 h** & **8 h** & **24 h** \\ \hline \multicolumn{2}{c}{**Apoptotic genes**} \\ \hline _Fas_ & \(145.0\pm 10.2\) & \(308.0\pm 5.7\) & \(146.0\pm 6.4\) & \(7.0\pm 0.9\) & \(32.0\pm 2.7\) & \(33.0\pm 8.7\) \\ \hline _FasL_ & \(26.0\pm 2.3\) & \(319.0\pm 8.7\) & \(88.0\pm 12.5\) & \(10.0\pm 1.4\) & \(27\pm 5.4\) & \(5.0\pm 1.8\) \\ \hline _BNP3_ & \(0.15\) & \(57.0\pm 6.7\) & \(5.0\pm 2.6\) & \(7.0\pm 2.3\) & \(16.0\pm 4.7\) & \(0.22\) \\ \hline _Casp3_ & \(1.8\pm 0.2\) & \(28.0\pm 5.2\) & \(19.0\pm 6.9\) & \(0.009\) & \(0.02\) & \(1.1\pm 0.1\) \\ \hline _Casp8_ & \(2794\pm 15.7\) & \(1285.0\pm 51.9\) & \(37.0\pm 4.2\) & \(42.0\pm 5.2\) & \(7.0\pm 4.9\) & \(3.0\pm 0.7\) \\ \hline _Cyt-C_ & \(3712\pm 20.8\) & \(5036.0\pm 44.2\) & \(1009.0\pm 10.4\) & \(4.0\pm 1.4\) & \(772.0\pm 13.8\) & \(2688.0\pm 17.5\) \\ \hline _Bax_ & \(0.01\) & \(0.07\) & \(2.0\pm 0.5\) & \(2.0\pm 0.7\) & \(5.0\pm 1.2\) & \(0.05\) \\ \hline _Bcl-2_ & \(10\pm 1.7\) & \(8.0\pm 2.8\) & \(3.0\pm 0.4\) & \(5.0\pm 1.8\) & \(3.0\pm 1.1\) & \(0.36\) \\ \hline _Bux/R0-2 ratio_ & \(0.01\) & \(0.01\) & \(0.7\) & \(0.01\) & \(0.7\) & \(13.9\pm 1.4\) \\ \hline _ATG5_ & \(0.45\) & \(1.1\pm 0.1\) & \(6.0\pm 1.7\) & \(0.14\) & \(0.88\) & \(1.5\pm 0.4\) \\ \hline _P53_ & \(0.45\) & \(0.13\) & \(0.53\) & \(0.06\) & \(0.81\) & \(2.0\pm 0.6\) \\ \hline \multicolumn{2}{c}{**Prolification genes**} \\ \hline _VEGF_ & \(0.45\) & \(0.52\) & \(0.03\) & \(3.0\pm 2.7\) & \(4.0\pm 1.3\) & \(0.03\) \\ \hline _TGF-\(\beta\)1_ & \(0.25\) & \(0.09\) & \(0.01\) & \(0.77\) & \(0.57\) & \(0.02\) \\ \hline _IGF1R_ & \(0.32\) & \(0.99\) & \(0.80\) & \(2.0\pm 0.7\) & \(1.3\pm 0.2\) & \(0.04\) \\ \hline _ATG12_ & \(0.20\) & \(0.28\) & \(0.22\) & \(0.12\) & \(0.36\) & \(0.03\) \\ \hline \multicolumn{2}{c}{**Onogenic genes**} \\ \hline _C-myc_ & \(0.16\) & \(0.15\) & \(0.03\) & \(0.18\) & \(0.29\) & \(0.05\) \\ \hline _TNF-\(\alpha\)_ & \(2.0\pm 0.5\) & \(4.0\pm 0.9\) & \(4.0\pm 0.7\) & \(0.02\) & \(0.04\) & \(0.01\) \\ \hline _NF-\(\alpha\)B_ & \(0.03\) & \(0.44\) & \(0.17\) & \(1.4\pm 0.2\) & \(2.6\pm 0.7\) & \(1.0\pm 0.1\) \\ \hline \multicolumn{2}{c}{**Kinase’s genes**} \\ \hline _PKC-\(\alpha\)_ & \(21.0\pm 8.7\) & \(2.0\pm 0.7\) & \(0.88\) & \(3.0\pm 0.9\) & \(2.0\pm 0.4\) & \(0.30\) \\ \hline _PRKAA1_ & \(0.49\) & \(42.0\pm 16.7\) & \(50.0\pm 7.8\) & \(0.77\) & \(0.5\) & \(7.0\pm 2.4\) \\ \hline _CDK-4_ & \(0.1\) & \(1.6\pm 0.3\) & \(0.47\) & \(9.0\pm 2.6\) & \(6.0\pm 1.5\) & \(0.1\) \\ \hline \hline \end{tabular}
\end{table}