A Functional Role For Pbr In Human Breast Cancer
Previous studies from this laboratory have shown that PBR plays a key role in steroidogenesis by mediating the translocation of cholesterol from the outer to the inner mitochondrial membrane . More recently, we have shown that PBR mediates cholesterol uptake even in nonmitochondrial membranes . To test whether PBR plays a similar role in MDA-231 nuclear membranes, intact nuclei were isolated from both MDA-231 and MCF-7 cells. Isolated nuclei were incubated with 10 nm cholesterol in the absence or presence of increasing concentrations of PK 11195 . In the absence of PK 11195, MDA-231 nuclear membranes demonstrated the ability to take up 30% more cholesterol relative to MCF-7 nuclei. In MDA-231 nuclei, addition of 108 to 106m PK 11195 resulted in a decrease in the amount of cholesterol retained by the nuclear membrane to levels relative to control treatment. This change corresponds to 0.52 pmol of cholesterol/mg of nuclear protein. MCF-7 nuclei failed to respond to the PK 11195 dose response . This change in cholesterol levels may reflect either reduced cholesterol uptake or increased release of nuclear membrane cholesterol. Considering our previous studies showing that perturbation of PBR by specific ligands induces the transport of cholesterol through bacterial membranes , we favor the second hypothesis.
Increased Expression Of The Pbr Correlates With Increased Aggressive Phenotype In Human Breast Cancer Cell Lines
Initial studies performed in MCF-7 human breast cancer cells indicated extremely limited expression of PBR . The data generated were insufficient for further Scatchard analysis. Before dismissing our original hypothesis, we decided to examine expression of the receptor in eight other human breast cancer cell lines using the PBR-specific, high-affinity ligand PK 11195. The results from these experiments indicated that those cell lines with a more invasive and chemotactic potential, such as HS-578-T and MDA-231, display dramatically increased levels of PBR binding relative to nonaggressive cell lines such as ZR-75-1, T47D, and MCF-7 . Furthermore, the MCF-7 Adriamycin-resistant derivative cell line, ADR, which expresses medium invasive and chemotactic potential as well as intermediate levels of CD44, expressed 2040% PBR binding relative to the MDA-231 cell line . Scatchard analysis of PBR binding in the MDA-231 and ADR cell lines further shows each to have a Bmax of 8.7 ± 1.4 and 1.3 ± 0.23 pmol/mg protein, respectively . As mentioned previously, despite obtaining specific PK 11195 binding in MCF-7 cells, the low binding levels were inadequate for estimating binding characteristics using Scatchard plot analysis .
Pbr Expression And Localization Correlates With Increased Aggressive Phenotype In Human Breast Tumor Biopsies
Before characterization of the receptor in the MDA-231 cells, we examined whether the differences seen in the expression and localization of PBR in human breast cancer cell lines reflects real differences found in human breast cancer or an artifact of the cell culture system. To this end, we examined the expression and localization of PBR in normal breast tissue procured by reductive mammoplasty and metastatic aggressive breast tumor biopsies. ,A shows HRP staining of the PBR antiserum used to detect the Mr 18,000 molecular weight protein in normal breast tissue. This section was counterstained with hematoxylin. A light immunostaining for PBR can be seen. The low staining is better seen in ,B in which the hematoxylin counterstain was omitted. These data were confirmed using a FITC-labeled secondary antibody . It should be noted that PBR appears to be localized in the ductal epithelium. ,D is the phase contrast image of the ,C. The level of expression and localization of the Mr 18,000 molecular weight PBR protein was further examined in normal human breast tissue using compound 4 . Incubation of the sections with unlabeled PK 11195 displaced the fluorescent labeling seen with compound 4 . Similarly, low levels of cytoplasmic expression were obtained in specimens from in situ and noninvasive breast tumors . These data were replicated in multiple sections taken from specimens from three separate individuals.
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Kodak Black Releases Limited Edition Tee For Breast Cancer Awareness Month
The month of October is known worldwide as National Breast Cancer Awareness Month. In honor of the annual international health campaign, Florida rapper Kodak Black has decided to show his support for breast cancer survivors by releasing a limited edition pink digital camo Sniper Gang t-shirt.
Now available in a wide range of sizes, the limited t-shirt can be purchased via the Sniper Gang Apparel website. Dropping in only one version, the tee features an all-around pink digital camo print with hits of white and black splattered throughout the camo aesthetic. Featuring Sniper Gangs signature logo on the front chest portion of the shirt, the tee runs true to size, and is limited in supply. Along with the purchase of the short sleeve military shirt, consumers will receive an exclusive pink Sniper Gang wristband to match with their shirts.
Kodak Blacks all-new digital camo pink tees are now available at snipergangapparel.com for a suggested price of $39.99. A portion of the proceeds from Kodaks Sniper Gang pink tees will go to the Florida Breast Cancer Foundation-floridabreastcancer.org.
Kodak Black Supports Breast Cancer
He a lil confused, but he got the spirit
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Determination Of Dna Content With Hoechst 33258
MDA-231 cells were plated on six-well plates at a concentration of 150,000 cells/well in DMEM supplemented with 10% FBS. After 24 h, cells were washed two times with PBS, and the medium was changed to DMEM supplemented with 0.1% FBS. After an additional 24 h, cells were treated with either 0, 1010, 108, 106, and 104m PK 11195 for 48 h. After incubation, cells were again washed two times with PBS and then solubilized with 200 l of 0.2% SDS in ETN buffer for 15 min at 37°C. Determination of DNA content was accomplished with the Hoechst 33258 dye as described previously .
Nuclear Transport Of Cholesterol
Nuclei were isolated from MDA-231 and MCF-7 as described by Elango et al.. Isolated nuclei were resuspended in 1 ml of ice-cold PBS. Cholesterol uptake in MDA-231 and MCF-7 nuclei was examined in the presence of 6.7 nmcholesterol and 3 g of nuclear protein for 60 min at 37°C . Samples were then centrifuged at 500 × g for 30 min, and pellets were washed in 500 ml of ice-cold PBS. After a second centrifugation at 500 × g for 30 min, 200 l of 1.0 n NaOH was then added to the pellets and incubated overnight at 37°C. After incubation, 200 l of 1.0n HCl was added, and samples were mixed vigorously. Three ml of scintillation cocktail were then added, and radioactivity was measured by liquid scintillation spectroscopy using a Wallac 1409 Liquid Scintillation Counter.
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Kodaks Heart Is In The Right Place
We always need a devil’s advocate.
I for one am PRO breast cancer
Ive never understood these awareness movements. We all knew breast cancer was a thing. Now we all buy shirts to prove we know about the thing. How does any of that do anything about the thing itself?
It is supposed to make more people get checked for cancer, because the sooner it is diagnosed the greater the chances for the patient. Most people realize they’re sick when it is already too late. Another thing is hospitals and research get more donations I think.
Basically it is a reminder about the disease.
Breast cancer is one if top three of not first in cancers in regards to awareness, Yet people truly arent aware of SARCOMA / BONE wich gets no Acknowledgement support etc. From Corp America These fake arse Celebs Sports Entertainment etc.
It’s time Support and awareness in made . Sarcoma Bone cancer YELLOW / RIBBON .. rare and deadly.
Iam stage two Survivor
Awareness movements can be a great thing, when there is little awareness of a cause of disease like the ALS Ice Bucket Challenge raising a lot of money for charity and promoting awareness of the disease. As you said, were all aware of breast cancer and we dont need to buy anymore pink shit because were all pretty informed on it.
Kodak Black And Supa Cindy Make Fans Proud They Host Event For Breast Cancer Patients/survivors
Kodak Black and Supa Cindy hosted an event for breast cancer patients and survivors. The fans could not be prouder, and they made sure to show this online as well.
TSR continued and noted that Kodak spoke about the women at the event, saying, ‘I know theyll appreciate it. I want to show these women theyre queens, theyre strong, and theyre going to get through it. Pray with them. Were going to get through it together.’
TSR noted that Taressa Hall, who is currently fighting breast cancer, said the event meant a lot to her.
‘It makes me continue to fight. It encourages me to go through this journey, continue through this journey, and it encourages me to be a witness to other people that we all can survive,’ said Taressa.
Somoene said: ‘One thing about Kodak, hes most definitely a philanthropist and I love to see it,’ and a commenter said: ‘Proud of Kodak for giving back. A lot of people dont use their access to give back and help others. Salute.’
A commenter posted this: ‘good he doing sum good w/ his money!’ and a follower said: ‘Kodak is night and day with the people they love him one day hate him the next.’
One other follower posted: ‘That’s what I’m tryna tell ya’ll. Kodak is always tryna give back. Always tryna give back!’
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Rapper Kodak Black Hosts Event In Miami For Women Fighting Breast Cancer
MIAMI As we near the end of breast cancer awareness month, a couple local celebrities decided they wanted to honor women who have beaten the disease and those who are still fighting it.
On Tuesday, rapper Kodak Black and radio host Supa Cindy hosted an event at Ives Estates Park where more than 20 women were given wigs, a cash gift, and even a journal with a note from the rapper himself.
I know theyll appreciate it, said Kodak Black. Show these women theyre queens, theyre strong, theyre going to get through it. Pray with them, were going to get through it together.
It was a sentiment that was felt by the woman attending the event.
It means a lot to me, said Taressa Hall, who is currently fighting breast cancer. It makes me continue to fight. It encourages me to go through this journey, continue through this journey and it encourages me to be a witness to other people that we all can survive.
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Breast Cancer Awareness Month
Show these women, that they queens, theyre beautiful, theyre strong. They gonna get through it, and pray with them we gonna get through it together, he said.
Kapri gifted each woman a wig from a local manufacturer, flowers and a gift card.
Taressa Hall is a three-year breast cancer survivor and says she appreciates the attention survivors and fighters receive during the month of October
It makes me continue to fight it encourages me to go through this journey continue this journey, and it encourages me to be a witness to other people that we all can survive we all we get this together, Hall said.
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Ways To Support Breast Cancer Awareness
In addition to wearing the classic pink ribbon, there are other ways you can show your support for breast cancer awareness. Here are 10 that may appeal to the techie in you.
When most people think about the colors of fall, they think of oranges, yellows, reds, and browns. But pink has been making more of an appearance lately, thanks to National Breast Cancer Awareness Month celebrated annually during the month of October. For the past 26 years, NBCAM has aimed to educate and empower both men and women about the dangers of breast cancer and the precautions women can take by catching it early with yearly screenings.
According to BreastCancer.org, about one in eight women in the U.S. will develop some form of breast cancer in her lifetime. Thanks to research and advances in technology, we’ve come a long way in terms of awareness and treatment over the last quarter of the century.
Things are a lot different now than they were back in 1985 when NBCAM began. The Web has become a great resource for education, support, and communication. Women can now use it as a means to seek support from other patients and survivors around the world and to find information to supplement the materials they receive from doctors. It has also helped the NBCAM organization gain awareness and help spread its message about “breast health, early detection and survivorship.”
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Fluorescent Microscopy With The Fluorescent Pbr Ligand Compound 4
MA-10, MDA-231, MCF-7, and ADR cells were grown on glass coverslips as described previously . Cells were then washed twice with sterile PBS and incubated for 45 min with 1 m compound 4, a fluorescent derivative of the PBR ligand FGIN-27, with or without a competing PBR ligand, FGIN-27, at a concentration of 100 m. After the incubation period, the cells were washed with PBS and examined by fluorescent microscopy using an Olympus BH-2 fluorescence microscope.
Analysis Of Human Biopsies
All human biopsies were obtained from the Lombardi Cancer Center Tumor Bank and the Department of Pathology at the Georgetown University Medical Center. Biopsies were analyzed and classified by the pathologists. Tumor biopsies were sectioned and then placed on glass slides. The slides were then heated at 56°C for 30 min. For specimens not in polyester wax, the slides were treated with xylene two times for 5 min each at room temperature. All slides were treated two times with 100% ethanol for 2 min each at room temperature and then two times with 95% ethanol for 2 min each at room temperature. After rinsing in running distilled H2O for 2 min, the slides were then treated with microwave fix in the microwave for 15 min at half-power and then 10 min at full power. The slides were then left to cool for 2 h at room temperature. For immunohistochemistry with anti-PBR primary antibodies, tissue sections were treated with a 30% H2O2:methanol mixture for 5 min at room temperature to neutralize endogenous peroxidase activity and then washed well with PBS. Primary antibody in 10% calf serum in PBS was added to sections at a concentration of 1:250 at room temperature for 1 h. Secondary antibody reactions using either HRP or FITC were carried out at concentrations of 1:500, as described above. For PBR fluorescent labeling with compound 4 , sections were treated with 1 × 107m compound 4 for 2 h at room temperature in a dark room and then viewed on an Olympus BH-2 fluorescent microscope.
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Kodak Black Donates Money & Gifts To Breast Cancer Survivors: ‘they’re Queens’
Northeast Miami-Dade, FL With Breast Cancer Awareness Month coming to a close, Kodak Black spent his Tuesday giving back in the Northeast Miami-Dade area as he showered breast cancer survivors with gifts and celebrated their journey and fight.
Kodak Black teamed up with 99JAMZ radio host Supa Cindy and met with more than 20 women to hand out wigs, cash, a personal journal and a note from Kodak himself.
I know theyll appreciate it, Kodak Black told WPLG. Show these women theyre queens, theyre strong, theyre going to get through it. Pray with them, were going to get through it together.
Cali Crip Cmac Says F Kodak Black & Sick Fetish For His Own Momma
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