{"id":10729,"date":"2016-12-22T10:50:34","date_gmt":"2016-12-22T10:50:34","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=10729"},"modified":"2020-04-24T09:35:38","modified_gmt":"2020-04-24T09:35:38","slug":"inhibition-of-fibrosarcoma-growth-by-5-hydroxy-7-ethoxy-flavanons-from-kaempferia-pandurata-roxb","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol9no3\/inhibition-of-fibrosarcoma-growth-by-5-hydroxy-7-ethoxy-flavanons-from-kaempferia-pandurata-roxb\/","title":{"rendered":"Inhibition of Fibrosarcoma Growth by 5-hydroxy-7-ethoxy-flavanons from Kaempferia pandurata Roxb"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

As a traditional medicine, the rhizome of Kaempferia pandurata Roxb is used as a medicine for dry cough, ulcers, disorders of the colon, stomach swelling, inflammation of the uterus, diare, and cancer (Hembing , 1994).<\/p>\n

The rhizome of Kaempferia pandurata Roxb extracted with diethyl ether produces relatively a large amount of 5-hydroxy-7-ethoxy-flavanones i.e. about 2,5% and alpinetin which is approximately 1%. 5-hydroxy-7-ethoxy-flavanones shows an antioxidants activity and relaxes smooth muscle. The high 5-hydroxy-7-ethoxy-flavanones content in the Kaempferia pandurata Roxb rhizome causes the isolation of the compound as a pure substance can be carried out easily and quickly. The 5-hydroxy-7-ethoxy-flavanones structure that the polarity reduces as a result of the existence of intramolecular hydrogen bonding between the carbonyl group at C-4 with a hydroxyl group at C-5, could be extractied with the application of semipolar or nonpolar solvents such as ethylacetate and CCl4<\/sub> (Parwata,1998). 5-hydroxy-7-ethoxy-flavanones shows a fairly high toxicity with LC50<\/sub> = 63.5 ppm (Parwata,2000). This compounds have an activity in inhibiting the growth of human breast cancer cells through the barrier activity of DNA topoisomerase I enzyme (Sukardiman et.al.,2006).<\/p>\n

The death of the DNA of a cancer cells can affect the the process inside the cell, especially the process of cell replication and terminated with the death of cancer cells by apoptosis (Sukardiman, et.al.,2006). DNA damage cells is thought to activate tumor suppressor gene p53 besides inducing \u00a0apoptosis (Whibley, 2009).<\/p>\n

Activation and stabilization of p53 products by a flavonoid, is thought to inhibit the angiogenesis effect through the mechanism of down regulation by the decrease of VEGF expression, COX-2 \u00a0and MMP-9 that involve in the metastasis process of cancer cells.. Based on this phenomenon the researchers suspect that 5-hydroxy-7-ethoxy-flavanones compound has the same activity as a therapy or chemopreventive to cancer on fibrosarkoma.<\/p>\n

Materials and Methods<\/strong><\/p>\n

The materials used were as follows: rhizome Kaempferia pandurata Roxb powder obtained from Badung Market, Denpasar-Bali and male mice of 2 months old and weight of 20-25 grams. The chemicals used were: technical n-heksana, n-heksana (E Merck), methanol (E Merck), ethanol (E Merck), chloroform (E Merck), ethyl acetat (E Merck) aquadest, silica gel and Plate of Thin Layer Chromatography (TLC)\u00a0 Aluminium\u00a0 with silica Gel GF-254, E Merck), primary and secondary antibodies p53 and VEGF, buffered formalin 10 %, PBS, DAB, H2<\/sub>O2<\/sub> 3% and destilate water (DW).<\/p>\n

The apparatus used were HPLC (High Peformance Liquid Cromatography) and TLC (Thin Layer Cromatography)-Densitometer Shimadzu CS-930, UV-Vis Spectrophotometer Hitachi 557 and 365 NIR Shimadzu, IR Spectrophotometer Jasco 5300 FTIR, proton NMR spectrometers and Carbon NMR Hitachi FT-1500 and Mass Spectrometer HP 8890, light microscope, polilysin preparate and objects glass<\/p>\n

Isolation and identification of 5-hydroxy-7-ethoxy-flavanones from the rhizome of Kaempferia pandurata Roxb<\/strong><\/p>\n

Approximately 2000 grams of \u00a0the powder of Kaempferia pandurata Roxb rhizome, was macerated with n-hexane for 24 hours, the liquid maserat obtained was then evaporated by rotary evavorator. The n- hexane extract was allowed to arise crystalline amorphous. After that, the amorphous crystals was chromatographed with vacuum column with n- hexane as the developer. \u00a0The results of this column chromatography were collected and then reevaporated with rotary evavorator until the crystals appeared. The crystals were filtered, then recrystallized with methanol 3 times. The crystals obtained in this study were shiny white. In the next step, the melting point of the crystals was determined by DTA and Melting Points Apparatus Fiesher & John and then its purity was tested by TLC (Thin Layer Chromatography) with the application of some eluents of various polarities, TLC-densitometer with multiple wavelengths as well as HPLC (High Performance Liquid Chromatography). When the test shows that 5-hydroxy-7-ethoxy-flavanones was pure, \u00a0the work was continued with subsequent identification by UV- Vis spectroscopy with sliding reagents, IR, 13<\/sup>C and 1<\/sup>H NMR, \u00a0MS and three-dimensional X-ray diffraction to find out the structure of 5-hydroxy-7-ethoxy-flavanones.<\/p>\n

Establishment of Fibrosarkoma on Mice by Benzopyrene Induction<\/strong><\/p>\n

Forty male mice were prepared to undergo a process of the research adaptation, then the mice had benzopyrene induction as much as 0.3 mg\/0.2 mL in the oleum olivarium as a subcutaneous injection in the scapular area 5 times every two days. All mice were maintained in the same atmosphere and diet for two months\/until the cancer formed in the neck. After the cancer volume reached \u00b1 100 mm3<\/sup>, the mice with cancer were randomly divided into 3 groups. Group I was the negative control which was only given CMC-Na. The second group was given 5-hydroxy-7-ethoxy-flavanones at a dose of 80 mg\/kg; group III was given cyclophosphamide at a dose of 13.33 mg\/kg. All of the test materials were administered orally and were given every day for 14 days. After that the mice were surgered for removing the fibrosarcoma tissue followed by weighing the tissue obtained.<\/p>\n

Results And Discussion<\/strong><\/p>\n

Result<\/strong><\/p>\n

Isolation<\/strong> \u00a0of 5-hydroxy-7-ethoxy-flavanones<\/strong><\/p>\n

Isolates obtained was in the form of needles white crystalline. Analysis of purity was performed by TLC using the mobile phase that different polarity. The results obtained showed one stain as shown in Table 1 below :<\/p>\n

Table 1: Analysis of purity isolates by Thin Layer Chromatography<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n
No.<\/strong><\/td>\nMobile phase<\/strong><\/td>\nResult<\/strong><\/td>\n<\/tr>\n
1<\/td>\nBAW = 4:1:5<\/td>\n1 stain<\/td>\n<\/tr>\n
2<\/td>\nCHCl3<\/sub>:CH3<\/sub>CH2<\/sub>OH=3:1<\/td>\n1 stain<\/td>\n<\/tr>\n
3<\/td>\nCHCl3<\/sub>:CH3<\/sub>CH2<\/sub>OH=15:1<\/td>\n1 stain<\/td>\n<\/tr>\n
4<\/td>\nnCH3<\/sub>CH2<\/sub>CH2<\/sub>CH2<\/sub>CH2<\/sub>CH3<\/sub>:CH3<\/sub>COOCH2<\/sub>CH3<\/sub> =1:1<\/td>\n1 stain<\/td>\n<\/tr>\n
5<\/td>\nnCH3<\/sub>CH2<\/sub>CH2<\/sub>CH2<\/sub>CH2<\/sub>CH3<\/sub>:CH3<\/sub>COOCH2<\/sub>CH3 <\/sub>=3:1<\/td>\n1 stain<\/td>\n<\/tr>\n
6<\/td>\nnCH3<\/sub>CH2<\/sub>CH2<\/sub>CH2<\/sub>CH2<\/sub>CH3\u00a0 <\/sub>: CH3<\/sub>COOCH2<\/sub>CH3<\/sub>= 3 : 2<\/td>\n1 stain<\/td>\n<\/tr>\n
7<\/td>\nCH3<\/sub>CH2<\/sub>CH2<\/sub>CH2<\/sub>CH2<\/sub>CH3\u00a0 <\/sub>: CH3<\/sub>COOCH2<\/sub>CH3<\/sub> = 7: 3<\/td>\n1 stain<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Note : BAW : n-Butanol : Acetic acid : Water All the Mobile phase result one stain \u00a0Analysis of \u00a0purity by TLC-densitometry at a wavelength [220 nm (I); 254 nm (II); 287 nm (III); 300 nm (IV); 320 nm (V) and 350 nm (VI)] resulted in one peak whereas by HPLC the purity of the compound obtained was of 94.83%. All this purity test results showed that the isolate was pure chromatographic Analysis of the melting point showed that the range of its melting point = 99-100o<\/sup>C (I); 99,5-101.5o<\/sup>C (II) and 99.5-100.5o<\/sup> C (III), \u00a0whereas with DTA melting point observed was of \u00a097o<\/sup>C, \u00a0and the substance stability was up to a temperature of 150o<\/sup>C. These results indicated that the isolates were pure based on its physical properties. Analysis of chemical constituents showed that isolates indicated positive flavonoids of flavanones, as shown by the following Table 2 :<\/p>\n

Table 2: Kualitatif Chemical Ingredients of isolates<\/strong><\/p>\n\n\n\n\n\n\n
No.<\/strong><\/td>\nReagent<\/strong><\/td>\nDiscoloration<\/strong><\/td>\nIndicated<\/strong><\/td>\n<\/tr>\n
1.<\/td>\nWillstater<\/td>\nColorless to pink colour<\/td>\n(+) Flavonoid<\/td>\n<\/tr>\n
2.<\/td>\nNaOH 10%<\/td>\nColorless to pink colour<\/td>\n(+) Flavonoid<\/td>\n<\/tr>\n
3.<\/td>\nFeCl3<\/sub><\/td>\nColorless to pink colour<\/td>\n(+) Flavonoid<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Note : All the Reagent result flavonoid<\/p>\n

Identification with UV-Vis spectrophotometer showed the l max = 287 nm (band II) and shoulder = 325 nm (band I).<\/p>\n

Subsequent analysis when isolates in methanol was coupled with AlCl3<\/sub>, the absorption band II has a bathochromic shift of 23 nm. Identification by IR spectroscopy showed some peaks as shown by the data in the following Table 3 :<\/p>\n

Table 3: Identification isolates by IR spectroscopy<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n
No.<\/strong><\/td>\nWavenumber (cm-1<\/sup>)<\/strong><\/td>\nIndicated<\/strong><\/td>\n<\/tr>\n
1<\/td>\n3437.46<\/td>\n-OH at C-5<\/td>\n<\/tr>\n
2<\/td>\n3061.31<\/td>\n-C-H stretch of aromatics<\/td>\n<\/tr>\n
3<\/td>\n1645.43<\/td>\n-C=O ketones at C-4<\/td>\n<\/tr>\n
4<\/td>\n1622.26 and 1579.64<\/td>\n-C=C- of aromatic<\/td>\n<\/tr>\n
5<\/td>\n3000-2800<\/td>\n-C-H of methyl<\/td>\n<\/tr>\n
6<\/td>\n1450-1350<\/td>\n-C-H of methylene<\/td>\n<\/tr>\n
7<\/td>\n1250 and 1290<\/td>\n-C-O<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Identification Isolates by Proton NMR spectroscopy resulted some peaks as shown by the data in the following Table 4 :<\/p>\n

Table 4: Identification Isolates by Proton NMR \u00a0spectroscopy<\/strong><\/p>\n\n\n\n\n\n\n\n\n
No.<\/strong><\/td>\nChemical shift (<\/strong>d<\/strong>), ppm<\/strong><\/td>\nIndicated<\/strong><\/td>\n<\/tr>\n
1<\/td>\n3.75 (s)<\/td>\nArOCH3<\/sub> (8H)<\/td>\n<\/tr>\n
2<\/td>\n6.019 (s)<\/td>\nH-6 and H-8 (2H)<\/td>\n<\/tr>\n
3<\/td>\n11.948 (s)<\/td>\n-OH (1H)<\/td>\n<\/tr>\n
4<\/td>\n2.817 to 3.029 (m)<\/td>\nH-3 cis or trans (2H)<\/td>\n<\/tr>\n
5<\/td>\n5,284<\/td>\nH-2 (1H)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Identification Isolates by Carbon NMR spectroscopy resulted some peaks as shown by the data in the following Table 5 :<\/p>\n

Table 5: Identification Isolates by Karbon \u00a0NMR \u00a0spectroscopy<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
No.<\/strong><\/td>\nChemical shift (<\/strong>d<\/strong>), ppm<\/strong><\/td>\nIndicated<\/strong><\/td>\n<\/tr>\n
1<\/td>\n55.583<\/td>\nO-CH3<\/sub> (1C)<\/td>\n<\/tr>\n
2<\/td>\n79.121<\/td>\nC-2 (1C)<\/td>\n<\/tr>\n
3<\/td>\n43.326<\/td>\nC-3 (1C)<\/td>\n<\/tr>\n
4<\/td>\n195.440<\/td>\nC-4 (1C)<\/td>\n<\/tr>\n
5<\/td>\n163.944<\/td>\nC-5 (1C)<\/td>\n<\/tr>\n
6<\/td>\n95.037<\/td>\nC-6 (1C)<\/td>\n<\/tr>\n
7<\/td>\n167.755<\/td>\nC-7 (1C)<\/td>\n<\/tr>\n
8<\/td>\n94.153<\/td>\nC-8 (1C)<\/td>\n<\/tr>\n
9<\/td>\n162, 572<\/td>\nC-9 (1C)<\/td>\n<\/tr>\n
10<\/td>\n103.025<\/td>\nC-10 (1C)<\/td>\n<\/tr>\n
11<\/td>\n138.241<\/td>\nC1 ‘(1C)<\/td>\n<\/tr>\n
12<\/td>\n138.241<\/td>\nC2’and C6 ‘(2C)<\/td>\n<\/tr>\n
13<\/td>\n128.667<\/td>\nC3′-C5 ‘(3C)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Analysis of isolates by mass spectroscopy resulted in molecular ion with m\/e=270 and the fragmentation as shown by the data in the following Table 6 :<\/p>\n

Table 6: Identification Isolates by Massa \u00a0spectroscopy<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
No.<\/strong><\/td>\nm\/e<\/td>\nIndicated<\/strong><\/td>\n<\/tr>\n
1<\/td>\n270<\/td>\nM+<\/sup><\/td>\n<\/tr>\n
2<\/td>\n252<\/td>\nM +<\/sup>– H2<\/sub>O<\/td>\n<\/tr>\n
3<\/td>\n242<\/td>\nM+`<\/sup>-CO<\/td>\n<\/tr>\n
4<\/td>\n227<\/td>\nM +<\/sup>-CO -CH3<\/sub><\/td>\n<\/tr>\n
5<\/td>\n213<\/td>\nM +<\/sup>-CO- C2<\/sub>H5<\/sub><\/td>\n<\/tr>\n
6<\/td>\n193<\/td>\nrelease of ring B\/phenyl<\/td>\n<\/tr>\n
7<\/td>\n166<\/td>\nM+<\/sup>-C2<\/sub>H5<\/sub>-C6<\/sub>H5<\/sub><\/td>\n<\/tr>\n
8<\/td>\n152<\/td>\nM+<\/sup> -C2<\/sub>H5<\/sub>-C6<\/sub>H5<\/sub>-CH3<\/sub><\/td>\n<\/tr>\n
9<\/td>\n123<\/td>\nM+<\/sup>-C2<\/sub>H5<\/sub>-C6<\/sub>H5<\/sub>-CO-CH3<\/sub><\/td>\n<\/tr>\n
10<\/td>\n77<\/td>\nC6<\/sub>H5<\/sub><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Note : M+ <\/sup>indicated Massa Molecole Relatif of 5-hydroxy-7-ethoxy-flavanones<\/p>\n

Surgery of fibrosarcoma<\/strong><\/p>\n

After one month of benzopyrene injection, fibrosarcoma started to grow and approximately after 2 months the volume of fibrosarcoma was 100 mm3<\/sup>. Prior to the treatment, one of the infected mice was sacrificed to prove the existence of fibrosarcoma. Justification of this was carried out by observing the histopathological tissue of the fibrosarcoma with HE colouring. The weight of the surgical result obtained was 3.4756 grams and the histopatology observation with 400x magnification showed a lot of chromatin (polychromatin) proving \u00a0the damage of the cells due to fibrosarcoma as shown by the following After one month of benzopyrene injection, fibrosarcoma started to grow and approximately after 2 months the volume of fibrosarcoma was 100 mm3<\/sup>. Prior to the treatment, one of the infected mice was sacrificed to prove the existence of fibrosarcoma. Justification of this was carried out by observing the histopathological tissue of the fibrosarcoma with HE colouring. The weight of the surgical result obtained was 3.4756 grams and the histopatology observation with 400x magnification showed a lot of chromatin (polychromatin), as shown by the following figure 6<\/p>\n\n\n\n
\"Figure<\/td>\n\n

Figure 1: Histopathology of fibrosarcoma tissue<\/strong><\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

(Note : arrows in the figure showed hole or chromatin)<\/p>\n

After 14 days of treatment, the mice with fibrosarcoma were sacrificed to take the cancer tissue for subsequent examination i.e. the weight of the cancer tissue, as shown by the data in the following table :<\/p>\n

Table 7: The Weight of Fibrosarkoma Mice after Treatment<\/strong><\/p>\n\n\n\n\n\n\n\n\n
Type of treatment<\/strong>\u00a0<\/strong><\/td>\nFibrosarkoma Weight (grams)<\/strong><\/td>\n<\/tr>\n
I<\/strong><\/td>\nII<\/strong><\/td>\nIII<\/strong><\/td>\nIV<\/strong><\/td>\nAverage<\/td>\n<\/tr>\n
without treatment<\/td>\n3.4756<\/td>\n3.4557<\/td>\n<\/td>\n<\/td>\n3.4656\u00b10.01<\/td>\n<\/tr>\n
CMC-Na\u00a0 [ Control (-)]<\/td>\n4.2765<\/td>\n4.1665<\/td>\n4.1095<\/td>\n4.1383<\/td>\n4.1727\u00b10.07<\/td>\n<\/tr>\n
5-hydroxy-7-ethoxy-flavanones 80 mm\/kg BW<\/td>\n1.0665<\/td>\n1.0605<\/td>\n1.1198<\/td>\n1.1027<\/td>\n1.0874\u00b10.02<\/td>\n<\/tr>\n
Cyclophosphamide [ Control ( + ) ]<\/td>\n0.1709<\/td>\n0.1021<\/td>\n0.2553<\/td>\n0.1021<\/td>\n0.1576\u00b10.07<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"Figure<\/td>\n\n

Figure 2: The Effect of\u00a0 5-hydroxy-7-ethoxy-flavanones on Fibrosarcoma Weight<\/strong><\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Discussion<\/strong><\/p>\n

Isolates obtained was in the form of needles white crystalline. Purity analysis with chromatographic by TLC , TLC – densitometry and HPLC showed one peak with a stain and a different polarity eluent. This shows that all this purity test results showed that the isolate was pure chromatographic. Analysis of one of the physical properties of the isolates are melting with Melting Points Apparatus and DTA showed that isolates said to be purely due to their melting point range < 2o<\/sup>. Based on these results further isolates identified spectroscopically. Phytochemical screening reagents Wilstatter color, FeCl3<\/sub> and NaOH 10% indicated that the isolates do indeed flavonoids flavanones.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Identification with UV-Vis spectrophotometer showed the l max = 287 nm (band II) and shoulder = 325 nm (band I). This was in agreement to the literature in which flavonoids flavanones show two absorption bands i.e. between 275-295 nm (band II) and a shoulder at 300-330 nm that caused by the cinnamoyl and benzoyl groups of flavanones as shown in the structure below (Silverstein et al , 1981; Parwata, 1998):<\/p>\n\n\n\n
\"Figure<\/td>\n\n

Figure 3: Structure of flavanone<\/strong><\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Bathochromic shift indicates that flavonoids flavanones contains an -OH group at C-5 as shown in the structure below (Silverstein et.al.,1981):<\/p>\n\n\n\n
\"Figure<\/td>\n\n

Figure 4: Structure of 5-hydroxy-7-ethoxy-flavanones<\/strong><\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Identification by IR spectroscopy proved that isolates containing -OH groups, -CH, -C=O and -C-O<\/p>\n

The results of the Proton and Carbon NMR spectra interpretation showed that 5-hydroxy-7-ethoxy-flavanones has 14 H atoms and 16 C, as shown by the following structure :<\/p>\n\n\n\n
\"Figure<\/td>\n\n

Figure 5: Numbering Structure of 5-hydroxy-7-ethoxy-flavanones<\/strong><\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Analysis with mass spectroscopy resulted in molecular ion with m\/e =270 that indicated the relative molecular mass of 5-hydroxy-7-ethoxy-flavanones with some fragmentation. This is illustrated by the following fragmentation schemes (Silverstein et al , 1981)<\/p>\n\n\n\n
\"Figure<\/td>\n\n

Figure 6: Pragmentasi of 5-hydroxy-7-ethoxy-flavanones<\/strong><\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

The result of the three-dimensional X-ray diffraction analysis showed that isolates as 5-hydroxy-7-ethoxy-flavanones have a structure as follows:<\/p>\n\n\n\n
\"Figure<\/td>\n\n

Figure 7: 5-hydroxy-7-ethoxy-flavanones structure with three-dimensional X-ray diffraction<\/strong><\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0<\/strong><\/p>\n

The weight of the surgical result obtained was 3.4756 grams and the histopatology observation with 400x magnification showed a lot of chromatin (polychromatin). proving \u00a0the damage of the cells due to fibrosarcoma<\/p>\n

After 14 days of treatment, the mice with fibrosarcoma were sacrificed to take the cancer tissue for subsequent examination i.e. the weight of the cancer tissue. The result obtained was that oral 5-hydroxy-7-ethoxy-flavanones concentration as much as of 80 mg\/kg body weight could inhibit the growth of fibrosarcoma with a weight of 68.62% grams while with the cancer drug (control +) there was a barrier of 95.95% compared to the negative control which was only given CMC-Na orally. This suggested that 5-hydroxy-7-ethoxy-flavanones is potentially be developed as a cancer chemotherapeutic agent.<\/p>\n

Conclusion<\/strong><\/p>\n

Based on the results of the research, it can be concluded as follows :<\/p>\n

Isolates produced are correct is 5-hydroxy-7-ethoxy-flavanones<\/p>\n

Results of histopathological analysis fibrosarcoma cells with HE staining showed there are many chromatin on fibrosarcoma as evidence of the destruction of normal cells due to the occurrence of fibrosarcoma<\/p>\n

5-hydroxy-7-ethoxy-flavanones has a significant effect in inhibiting the growth of fibrosarcoma.<\/p>\n

5-hydroxy-7-ethoxy-flavanones oral concentration of 80 mg\/kg body weight could reduce as much as 68.62 % fibrosarcoma weight while with a cancer drug (control +) there was adecrease of 95.95 % , this suggested that 5-hydroxy-7-ethoxy-flavanones is potentially be developed as a cancer chemotherapy agent<\/p>\n

Suggestion<\/strong><\/p>\n

This research should be continued on the effect of the treatment of p53, COX-1, COX-2, VEGF and MMP-9<\/p>\n

Acknowledgement<\/strong>s<\/strong><\/p>\n

The author would like to thank profusely all parties, especially Mr. Eko Adiputra for his help in the maintenance and surgery of the mice in the Animals Laboratory, Faculty of. Pharmacy-Airlangga University during the research. We also thank profusely the Chief and Staff of LPPM Udayana University for providing the funds to afford this research.<\/p>\n

References<\/strong><\/p>\n

    \n
  1. Adi Parwata, 1998, Isolasi, Identifikasi Senyawa 5-hydroxy-7-ethoxy-flavanones pada\u00a0Rimpang Temu Kunci (Kaempferia pandurata Roxb) dan Standarisasi Ekstrak Etanol berdasarkan Kadar 5-hydroxy-7-ethoxy-flavanonesnya dengan KLT Densitometri.\u00a0\u00a0<\/em>Thesis, <\/em>PPS Unair,\u00a0 Surabaya.<\/li>\n
  2. Adi Parwata, 2000,Uji Toksisitas Senyawa 5-hydroxy-7-ethoxy-flavanones pada Rimpag Temu Kunci (Kaempferia pandurata Roxb), DIK\/DIKS, Lemlit Unud, Bali.<\/li>\n
  3. Anonim, 2005, Fibrosarkoma, http:\/\/medika-online.blogspot.com\/2005\/11\/\u00a0<\/u>Fibrosarkoma.html , diakses, 28 September 2011<\/li>\n
  4. Bail,L., Aubourg L., Habrioux G., 2000. Effect of 5-hydroxy-7-ethoxy-flavanones on <\/em>estrogen <\/em>metabolism and estrogen reseptor transactivivation<\/em>, Cancer Lett, Aug, 1;156910;37-44.<\/li>\n
  5. Banerjee S. , Carlos ,B.R and Bharat B.A.(2002) Suppression of 7,12-Dimethylbenz(a) anthracene-induced Mammary Carcinogenesis in Rats by Resveratrol , Carcinogenesis, 62, 4945-4954.<\/li>\n
  6. Cumming,J., Smyth,J.F.,1993. DNA Topoisomerase I and II as target of Rational Design of <\/em>New Anticancer Drugs<\/em>, Ann Oncology, Aug, 3(7), 533-534.<\/li>\n
  7. Harborne J.B. and Mabry, T.J., 1992, Flavonoids, Advances and Research, Chapman\u00a0and Hall, London.<\/li>\n
  8. Jianguo,M., Karin, Reed,A., and James,M., Gallo, 2002, Cells Designed to Deliver\u00a0<\/em>Anticancer Drug by Apoptosis<\/em>, Cancer Research, 62, 1382-7.<\/li>\n
  9. Manahan,D., and Wienberg,R.A., 2002. The Hallmarks of Cancer<\/em>, Cell, 100 : 57-70.\u00a0Markam K.R., 1988, Cara Identifikasi Flavonoid,terjemahanPenerbit \u00a0ITB Bandung,1-59<\/li>\n
  10. Nagata ,S. 1997. Apoptosis by Death Factor<\/em>, Cell. 88 : 355 \u2013 365.<\/li>\n
  11. Pai,S., Kaustubh,D. and Debabrata,M., 2001. Central Role of p53 Regulation of Vascular <\/em>Permeability Factor\/Vascular Endothelial Growth Factor (VPF\/VEGF) Expression in Mammary Carinoma,<\/em> Cancer Research, 61, 6952-6957.<\/li>\n
  12. Ravi,R.,Bijoyesh,M., Zaver M.B., Carrie. H.S., Dmitri,A., 2000. Regulation of tumor\u00a0<\/em>angiogenesis by p53-induced degradation of hypoxia-inducible factor 1\u03b1<\/em>, Gene and Development, Vol 14.<\/li>\n
  13. Silverstein, Bassler and Morrill, 1981, Spectrometric Identification of Organic\u00a0Compounds, John Willey and Sons, New York.<\/li>\n
  14. Sukardimanb<\/sup>, Darwanto A, Tanjung M., Parwarta\u00a0 Oka Adi ., 2000. Cytotoxic Mechanism of <\/em>\u00a0\u00a0\u00a0<\/em>Flavonoid from Temu Kunci (Kaempferia pandurata) in Cells Culture of Human Mammary Carcinoma<\/em>, Clinical Hemorheology and Microcirculation, 23 ,185-190.<\/li>\n
  15. Sukardiman, Noor Cholies Z, Sismindari ,2006. Induksi Apoptosis dan Peningkatan\u00a0 Ekspresi p53, Bax\u00a0 serta Aktivasi Enzim Caspase Sel Kanker Payudara Manusia oleh 5-hydroxy-7-ethoxy-flavanones dari<\/em> Kaempferia pandurata<\/em> Roxb, Laporan Penelitian Hibah Bersaing Tahun I, Lembaga Penelitian Universitas Airlangga.<\/li>\n
  16. Wittmann\u00a0S, Bali\u00a0P, Donapaty\u00a0S, Nimmanapalli\u00a0R, Guo\u00a0F, Yamaguchi\u00a0H, Huang\u00a0M, Jove\u00a0R, Wang\u00a0HG & Bhalla\u00a0K. (2003). Flavopiridol down-regulates antiapoptotic proteins and sensitizes human breast cancer cells to epothilone B-induced apoptosis<\/em>. Cancer Res 63: 93\u221299.<\/li>\n
  17. Whibley C., 2009, p53 polymorphisms: cancer implications, Nature RevIew\u00a0 Cancer, Vol. 9,\u00a0 Februari 2009, p. 95-107, http:\/\/www. google.co.id\u00a0\u00a0 \/p53 +cancer+search,Diakses,\u00a0 6 Desember 2011.<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    Introduction As a traditional medicine, the rhizome of Kaempferia pandurata […]<\/p>\n","protected":false},"author":8,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[35],"tags":[],"class_list":["post-10729","post","type-post","status-publish","format-standard","hentry","category-vol9no3"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/10729","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=10729"}],"version-history":[{"count":4,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/10729\/revisions"}],"predecessor-version":[{"id":32697,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/10729\/revisions\/32697"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=10729"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=10729"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=10729"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}