Cell And Breast Cancers And Colorectal Cancer

However, the majority of known anti-cancer drugs target normal cells as well. Methotrexate, formerly known as amethopterin, is an antimetabolite used in the treatment of some forms of cancer (Shacter and Law, 1956). It inhibits the function of dihydrofolate reductase (DHFR) by tightly binding to the enzyme. Following this slow interaction between methotrexate and DHFR, an enzyme-NADPH-inhibitor complex is formed (Stone et al., 1984). Therein, DHFR is rendered unable to catalyze the NADPH-dependent reduction of dihydrofolate to tetrahydrofolate (Anderson, 2017). Tetrahydrofolate is an important methyl group shuttle in the de novo synthesis of purine and pyrimidine nucleotides and some amino acids (Barbara and Hiroshi, 2002). Therefore, eliminating it from cells is a good therapeutic strategy against cancer cells, as it would limit the de novo synthesis of purines and pyrimidines (Lane and Fan, 2015). Inhibiting the function of DHFR by its competitive inhibitor methotrexate is one such good therapeutic strategy, as it would disable DHFR from converting dihydrofolate into tetrahydrofolate. However, some types of cancer cells had acquired some forms of resistance against methotrexate (Hans et al., 2011). Equally important, methotrexate should be administered in well established low doses, because it affects

In vitro: Daun02 was found to be a good substrate for β-gal, while gal-DCN4 was a poor substrate. Compared with mock-transduced control cells, except for PC3, the β-gal-transduced tumors showed 3- to 60-fold increased sensitivity to Daun02. In addition, in culture medium containing β-gal-transduced cell lines but not in the medium from mock-transduced controls, daunomycin was formed from Daun02 [1].

Picture 3. This picture shows the stock cultures for both colony 1 and colony 2. The left test tube shows the growth from colony 1 on the TSA slant. The right test tube shows the growth from colony 2 on the TSA slant.

After collecting all eight samples, they were then placed in the Petri dishes. The Petri dishes were both placed in an incubator. The temperature at that time was 80 degrees Fahrenheit. The samples remained in the incubator for over 24 hrs, by which time, quite a lot of growth was formed.

A 6-well plate containing pre-plated lung cancer cells (cell line H460) in 3mL of media was prepared for the purposes of this lab. Two wells were labeled NT (no treatment) and LPS (lipopolysaccharide). The well plate was tipped to pool the media to the base of the well. 30 μL of LPS was added to the LPS well, and swirled to mix its content. As the cell was incubated for 15 minutes, 225 μL of RIPA and 25 μL of protease inhibitor were added to a 1.5 mL tube, and kept on ice. Six 1.5 mL tubes were labeled NT-1, NT-2, NT-DIL, LPS-1, LPS-2 and LPS-DIL, and placed on ice. After 15 minutes all media was removed from the NT and LPS wells using the P1000 micropipette at 1000 μL. The remaining media was removed with the pipette by titling the plate allowing

Studies have shown that unlike insulin glargine, metformin has the potential to reduce both the cancer-causing instances of genetic mutation and the associated cellular damage (5). It can interfere with the growth of tumors both directly and indirectly (2); directly by inhibiting the growth of cancer cells,

After 10 minutes, place the tubes into a test tube float and put tubes into a water bath at 42 degrees Celsius for 50 seconds. After 50 seconds, immediately put the tubes back on ice for 2 minutes. Use a P-1000 micropipette to transfer 250 ul of LB nutrient broth to both tubes, making sure to use a new sterile pipette tip each time. Allow the cells to recover by incubating the tubes in a 37 degree Celsius water bath during 15 minutes. Use a pipette with a new sterile tip to transfer 200 ul of each onto the according plates. Use a new sterile inoculation loop; spread the solution using a disposable sterile L-shape across the agar plate. Stack the plates and tape them together. Place the stack upside down in a 37 degrees Celsius incubator foe 24-36 hours. After 24 hours, determine which plate is observed grown

In the 10-3 pasteurized sample, the plate exhibited 71,000 cells/mL. The results of the additional dilution samples contained too few colony forming units to count. However, in the 10-7 dilution, although the plate demonstrated 12 colonies, there should have been no colony forming units on this plate. The reasons for this could have been that this sample was contaminated from “double-dipping” the sample before dispensing it onto the plate or when using the pipette, it mistakenly was inserted in a higher concentration sample and then immediately to a lower concentration sample before it was dispensed onto the plate.

Chemotherapy is done to either cure cancer or to prolong life. The chemotherapeutic agents used in this process, acts by targeting cancer cells. In this process, they also harm the normal cells in the patient’s body, leading to adverse health effects. Most of the conventional chemotherapeutic agents available today, do not have the expected specificity, and can end up harming normal cells to varying

The data showed in figure 6, are form Boyden chamber assay that was used to observe the invasion or movement of cells from one place to another. In this assay the authors used three treatment groups; i. 1mM melatonin supplemented with 10% FBS (Fetal Bovine Serum), ii. 10% FBS supplement defined as positive control, and iii. 0.5% FBS supplement as a negative control. Although the figure was shown in two sub-part, but we can compare the melatonin group with the negative control group because all data were normalized by positive control. The authors conducted the Boyden chamber assay in canine (CMT-U229) and human (MCF-7) breast cancer stem cells. 1mM melatonin decreased cell invasion by 40.4% in canine breast cancer stem cells and by 56.3% in human breast cancer stem cells compared to the positive control. However, negative control further decreased cell invasion compared to melatonin treatment. Although the authors did not mention any exact number, it could be assumed from figure 6 that negative control decreased cell invasion by about 90% in canine breast cancer stem cells and by about 95% in human breast cancer stem cells compared to positive control. Therefore, the percentage of inhibition of cell invasion was higher for negative control compared to the melatonin group in both types of cancer stem cells. However, we cannot confirm that this comparison is precise due to lack of statistical comparison. Student’s t-test was used to compare between two groups, whereas, ANOVA needs to be used for three or more groups. Therefore, if we want to compare among positive control, melatonin treatment and negative control we need to run one-way ANOVA

The present study investigates the cytogenetic changes that could be induced in cancer cells after treatment with PDT. In this work, we evaluated the cytotoxic and genotoxic effects that could be induced by 5-ALA/PDT on two different cancer cell lines; adenocarcinoma breast cancer (MCF-7) and hepatocellular carcinoma (HepG2). When MCF-7 and HepG2 cell lines treated with 5-ALA (0.5, 1, 1.5 and 2mM) alone for 4h then incubated for 24h, 5-ALA did not induce cytotoxic effects on both cell lines. Also, laser irradiation alone did not induce a cytotoxic effect on MCF-7 or HepG2 cells after exposure for 4min then incubation for 24h. These results are in agreement with the observations of Chu et al. [21], who found that both 5-ALA and its

incubation at 28 ° C for 20, 40, 80, 160 and 350 days. After 350 days, 56% of 2,3,7,8-

This value aided in differentiating between cell losses due to the washes and cell losses due to cell mortality resulting from the mutagenesis. Next, number of cells in 4x4 grid was counted under a microscope from each members of the group. The average value happened to be 54.5 multiplied by 2,500,000 cells equaled to 136, 250, 000 cells/mL Experimental. To determine the cells/ mL plated experimental, number of colonies on the 0.1mL plate (23) count was multiplied by the dilution factor (10), which equaled to 230 cells/mL plated experimental. Then the cells/mL Experimental (136,250,000) was multiplied by the correction factor of 0.18 then divided by 100,000 dilution factor which resulted as 253.425 cells plated. The final step was taking the Experimental plate count of 230 cells/mL divided by cell plated of 253.425 multiplied by 100 which happened to be 90.7 percent of

The breast cancer cells were incubated at 37 °C under normal atmospheric pressure, while the normal breast cancer cells were incubated at 37 °C in environments with 5% CO2 levels, respectively. The MDA-MB-231 and Hs578bst cells were grown respectively in L-15 or 46-X base media. These were supplemented with 100 I.U./ml penicillin/100 μg/ml streptomycin and 10% FBS. The cells were first rinsed with 1X PBS and then fixed with 4 wt% paraformaldehyde solution for 10 mins at room temperature. They were then rinsed with PBS solution for three more times, before a final rinse with purified water. Finally, the samples were dried in a vacuum desiccator for at least 5 hours.

This assay was performed in 96 well plates at a cell density of 3x103 cells/well. Cancer cells precultured in media depend on type of