Abstract

Effect of Thai Traditional Herbal Crude Extracts on Cancer Cell Death

BUNTHITA JONGPREMKITPAISAN, SUWAPAK CHAIWASUKUN

ASST. PROF.PORNTIP WAIWUT, Ph.D.

CANCER, APOPTOSIS, THAI TRADITIONAL HERBS, DR5, XIAP

EFFECT OF THAI TRADITIONAL HERBAL EXTRACT ON CANCER CELL DEATH BUNTHITA JONGPREMKITPAISAN, SUWAPAK CHAIWASUKUN DOCTOR OF PHARMACY PHARMACEUTICAL SCIENCES ASST. PROF.PORNTIP WAIWUT, Ph.D. CANCER, APOPTOSIS, THAI TRADITIONAL HERBS, DR5, XIAP Chemotherapy is a main treatment for cancers but there are several side effects occur. Hence, searching for new active ingredients from plants for inhibition of cancer cell proliferation and less side effect was our interest. To investigate the cytotoxic effect and mechanism induced apoptosis on colon adenocarcinoma cell (HT-29) and hepatocellular carcinoma cell (HepG2) of Thai traditional herbal extracts including Cratoxylum formosum, Limnophila geoffrayi, Aegle marmelos, Syzygium gratum and Fagraea fragrans. The cytotoxic effect of the extract on cancer cell and normal cell were measured by MTT assay and the cell morphology was assessed by using phase contrast microscopy. The effect of extracts on apoptotic signaling protein was investigated by Western blot analysis. The result showed that Limnophila geoffrayi extract at 1 mg/ml had the highest cytotoxicity significantly different from a control group towards both HT-29 cell (p<0.001) and HepG2 cell (p<0.05) with IC50 were 0.57 ± 0.03 mg/ml and 0.57 +0.01 mg/ml respectively. Cratoxylum formosum extract showed cytotoxic effect on HT-29 cell at 1 mg/ml significantly (p<0.01) with IC50 was 0.87 + 0.03 mg/ml and Aegle marmelos extract showed cytotoxic effect on HepG2 at 1 mg/ml significantly (p<0.05) with IC50 was 0.76 + 0.02 mg/ml. In apoptosis pathway, Limnophila geoffrayi and Cratoxylum formosum extract induced cell death on HT-29 cell by decreasing level of X-linked inhibitor of apoptosis (XIAP) and inducing casepase-3 activation. In HepG2 cell, Limnophila geoffrayi and Aegle marmelos extract induced apoptosis through upregulation of DR5, decreasing XIAP level and inducing caspase-3 activation. Moreover, the three extracts showed no cytotoxic effect significantly towards normal cell . We found that Limnophila geoffrayi and Cratoxylum formosum extracts could inhibit cell proliferation on HT-29 cell through inducing apoptosis pathway by inhibition of XIAP and activation of caspase-3. Limnophila geoffrayi and Aegle marmelos extracts could inhibit cell proliferation on HepG2 cells by inducing apoptosis through inhibition of XIAP,upregulation of DR5 and caspase-3 activation. However, they had no harm to normal cell. EFFECT OF THAI TRADITIONAL HERBAL EXTRACT ON CANCER CELL DEATH BUNTHITA JONGPREMKITPAISAN, SUWAPAK CHAIWASUKUN DOCTOR OF PHARMACY PHARMACEUTICAL SCIENCES PROF.PORNTIP WAIWUT, Ph.D. CANCER, APOPTOSIS, THAI TRADITIONAL HERBS, DR5, XIAP Chemotherapy is a main treatment for cancers but there are several side effects occur. Hence, searching for new active ingredients from plants for inhibition of cancer cell proliferation and less side effect was our interest. To investigate the cytotoxic effect and mechanism induced apoptosis on colon adenocarcinoma cell (HT-29) and hepatocellular carcinoma cell (HepG2) of Thai traditional herbal extracts including Cratoxylum formosum, Limnophila geoffrayi, Aegle marmelos, Syzygium gratum and Fagraea fragrans. The cytotoxic effect of the extract on cancer cell and normal cell were measured by MTT assay and the cell morphology was assessed by using phase contrast microscopy. The effect of extracts on apoptotic signaling protein was investigated by Western blot analysis. The result showed that Limnophila geoffrayi extract at 1 mg/ml had the highest cytotoxicity significantly different from a control group towards both HT-29 cell (p<0.001) and HepG2 cell (p<0.05) with IC50 were 0.57 ± 0.03 mg/ml and 0.57 +0.01 mg/ml respectively. Cratoxylum formosum extract showed cytotoxic effect on HT-29 cell at 1 mg/ml significantly (p<0.01) with IC50 was 0.87 + 0.03 mg/ml and Aegle marmelos extract showed cytotoxic effect on HepG2 at 1 mg/ml significantly (p<0.05) with IC50 was 0.76 + 0.02 mg/ml. In apoptosis pathway, Limnophila geoffrayi and Cratoxylum formosum extract induced cell death on HT-29 cell by decreasing level of X-linked inhibitor of apoptosis (XIAP) and inducing casepase-3 activation. In HepG2 cell, Limnophila geoffrayi and Aegle marmelos extract induced apoptosis through upregulation of DR5, decreasing XIAP level and inducing caspase-3 activation. Moreover, the three extracts showed no cytotoxic effect significantly towards normal cell . We found that Limnophila geoffrayi and Cratoxylum formosum extracts could inhibit cell proliferation on HT-29 cell through inducing apoptosis pathway by inhibition of XIAP and activation of caspase-3. Limnophila geoffrayi and Aegle marmelos extracts could inhibit cell proliferation on HepG2 cells by inducing apoptosis through inhibition of XIAP,upregulation of DR5 and caspase-3 activation. However, they had no harm to normal cell. EFFECT OF THAI TRADITIONAL HERBAL EXTRACT ON CANCER CELL DEATH BUNTHITA JONGPREMKITPAISAN, SUWAPAK CHAIWASUKUN DOCTOR OF PHARMACY PHARMACEUTICAL SCIENCES ASST. PROF.PORNTIP WAIWUT, Ph.D. CANCER, APOPTOSIS, THAI TRADITIONAL HERBS, DR5, XIAP Chemotherapy is a main treatment for cancers but there are several side effects occur. Hence, searching for new active ingredients from plants for inhibition of cancer cell proliferation and less side effect was our interest. To investigate the cytotoxic effect and mechanism induced apoptosis on colon adenocarcinoma cell (HT-29) and hepatocellular carcinoma cell (HepG2) of Thai traditional herbal extracts including Cratoxylum formosum, Limnophila geoffrayi, Aegle marmelos, Syzygium gratum and Fagraea fragrans. The cytotoxic effect of the extract on cancer cell and normal cell were measured by MTT assay and the cell morphology was assessed by using phase contrast microscopy. The effect of extracts on apoptotic signaling protein was investigated by Western blot analysis. The result showed that Limnophila geoffrayi extract at 1 mg/ml had the highest cytotoxicity significantly different from a control group towards both HT-29 cell (p<0.001) and HepG2 cell (p<0.05) with IC50 were 0.57 ± 0.03 mg/ml and 0.57 +0.01 mg/ml respectively. Cratoxylum formosum extract showed cytotoxic effect on HT-29 cell at 1 mg/ml significantly (p<0.01) with IC50 was 0.87 + 0.03 mg/ml and Aegle marmelos extract showed cytotoxic effect on HepG2 at 1 mg/ml significantly (p<0.05) with IC50 was 0.76 + 0.02 mg/ml. In apoptosis pathway, Limnophila geoffrayi and Cratoxylum formosum extract induced cell death on HT-29 cell by decreasing level of X-linked inhibitor of apoptosis (XIAP) and inducing casepase-3 activation. In HepG2 cell, Limnophila geoffrayi and Aegle marmelos extract induced apoptosis through upregulation of DR5, decreasing XIAP level and inducing caspase-3 activation. Moreover, the three extracts showed no cytotoxic effect significantly towards normal cell . We found that Limnophila geoffrayi and Cratoxylum formosum extracts could inhibit cell proliferation on HT-29 cell through inducing apoptosis pathway by inhibition of XIAP and activation of caspase-3. Limnophila geoffrayi and Aegle marmelos extracts could inhibit cell proliferation on HepG2 cells by inducing apoptosis through inhibition of XIAP,upregulation of DR5 and caspase-3 activation. However, they had no harm to normal cell.