## 3 Results
### Effect of Gefitinib and Caneritinib on Total Expression and Activation of EGFR and HER-2 Proteins  
We further evaluated the effect of gefitinib and canertinib on the protein level of EGFR and HER-2 and their associated phosphorylation at tyrosine 1173 and tyrosine 1248 residues, respectively in ascitie fluid-derived cell samples. As shown in Table 4A, in the EGFR positive cells, five out of nine samples (patient numbers 1, 4, 6, 10, 20; \(p<0.05\)) showed a decrease in EGFR expression after gefitinib treatment. The phosphorylation of EGFR was also compromised by gefitinib. Six out of nine astictic cell samples (patient numbers 1, 3, 6, 10, 17, 20; \(p<0.05\)) showed a decrease in the phosphorylation of EGFR after gefitinib treatment. Also, canertinib reduced the level of EGFR and pEGFR. Canertinib reduced the total expression of EGFR in five out of nine cells samples (patient numbers 1, 3, 4, 6, 20; \(p<0.05\)) and pEGFR in five samples (patient numbers 1, 3, 4, 17, 20; \(p<0.05\)). \begin{table}
\begin{tabular}{c c c c} \hline \hline  & & \multicolumn{3}{c}{**(A) (+) EGFR**} \\ \hline \multirow{2}{*}{**Patient ID**} & \multirow{2}{*}{**Control**} & \multicolumn{3}{c}{**Cellular Viability**} \\  & & \multicolumn{3}{c}{**(Relative to Control \%)**} \\ \cline{3-4}  & & **5 \(\upmu\)[PERSON]** & **5 \(\upmu\)[PERSON]** \\ \hline \multicolumn{1}{c}{OVCAR-5} & 100 & \(99.9\pm 1.3\) & \(102\pm 1.2\) \\
1 & 100 & \(74.6\pm 2.7\,\ast\) & \(87.8\pm 17.6\) \\
3 & 100 & \(98.9\pm 6.7\) & \(104.5\pm 4.6\) \\
4 & 100 & \(84.6\pm 0.8\,\ast\) & \(74.9\pm 4.3\,\ast\)\# \\
6 & 100 & \(95.5\pm 3.1\) & \(91.7\pm 1.9\,\ast\) \\
8 & 100 & \(93.6\pm 3.1\) & \(92.5\pm 3.9\) \\
10 & 100 & \(102.8\pm 5.5\) & \(95.4\pm 2.7\) \\
17 & 100 & \(86.3\pm 1.1\,\ast\) & \(89.8\pm 1.3\,\ast\) \\
19 & 100 & \(86.9\pm 2.0\,\ast\) & \(78.4\pm 4.1\,\ast\) \\
20 & 100 & \(102.8\pm 5.5\) & \(95.4\pm 2.7\) \\ \hline \multicolumn{1}{c}{} & \multicolumn{3}{c}{**(B) (+) EGFR/HER-2**} \\ \hline \multirow{2}{*}{**Patient ID**} & \multirow{2}{*}{**Control**} & \multicolumn{3}{c}{**Cellular Viability**} \\  & & **(Relative to Control \%)**} \\ \cline{3-4}  & & **5 \(\upmu\)[PERSON]** & **5 \(\upmu\)[PERSON]** \\ \hline \multicolumn{1}{c}{SKOV-3} & 100 & \(95.2\pm 2.7\) & \(82.9\pm 2.8\,\ast\) \\
7 & 100 & \(105.1\pm 2.1\) & \(117.1\pm 4.5\) \\
9 & 100 & \(91.7\pm 30\) & \(81.4\pm 33.3\,\ast\) \\
11 & 100 & \(87.8\pm 6.5\) & \(82.8\pm 7.4\,\ast\) \\
12 & 100 & \(86.5\pm 6.8\) & \(90.3\pm 4.6\) \\
13 & 100 & \(94.3\pm 4.4\) & \(92.7\pm 4.3\) \\
15 & 100 & \(95.4\pm 3.1\) & \(93.9\pm 2.1\) \\
16 & 100 & \(95.6\pm 6.2\) & \(88.2\pm 1.9\,\ast\) \\
18 & 100 & \(99.8\pm 4.9\) & \(102.7\pm 2.5\) \\ \hline \multicolumn{1}{c}{} & \multicolumn{3}{c}{**(C) (+) EGFR/HER-2**} \\ \hline \multirow{2}{*}{Patient ID} & \multirow{2}{*}{**Control**} & \multicolumn{3}{c}{**Cellular Viability**} \\  & & **(Relative to Control \%)**} \\ \cline{3-4}  & & **5 \(\upmu\)M Gef** & **5 \(\upmu\)M Can** \\ \hline
2 & 100 & \(68.3\pm 0.11\,\ast\) & \(65.3\pm 11.2\,\ast\) \\
5 & 100 & \(95.3\pm 058\) & \(97.6\pm 0.78\) \\
14 & 100 & \(96.45\pm 1.4\) & \(96.56\pm 0.12\) \\ \hline \hline \end{tabular}
\end{table} 
Table 3: Cellular viability of OVCAR-5, SKOV-3 and astic derived cells from 20 patients are affected by gefitinib (Gef) and canertinib (Can). Nine patient cell lines and the OVCAR-5 cells expressing EGFR only, have a decrease in viability by the treatment of Gef and Can (**A**). Eight patient cell lines and SKOV-3 cells expressing both EGFR and HER-2 do not respond to Gef, but have increased responsiveness to Can (**B**). Only one cell sample of EGFR/HER-2 negative cells responded to both gefitinib and canertinib (**C**). Representative analysis was from a set of two independent experiments, and each experiment had three duplicates. Mean \(\pm\) SE; \(n=2\) is presented and statistical significance was determined by a \(t\)-test (\(\ast,p<0.05\)). The statistical difference between gefitinib and canertinib treated cells was determined by a \(t\)-test (\(\#\), \(p<0.05\)).