Point to be noted here is the androgen dependency of the cell lines. It is known that PC3 and DU145 are androgen independent and LNCaP is androgen dependent. How far is this androgen dependency responsible for the problem i.e. prostate cancer?
So first query here is, which cell line should be given preference keeping in view the androgen dependent or independent nature?
One of the causes of prostate cancer is an androgenic disorder which arises mainly by the ill effects of testosterone metabolism and its conversion to dihydrotestosterone in the presence of 5alpha reductase. Benign prostate hyperplasia (BPH) also has the same reason for its development in adult males.
So is it good to analyze the prostate cancer cell lines PC3 and DU145 or LNCaP also and then arrive at a conclusion about the effect of the sample analyzed?
Or should one go for the analysis of the 5-alpha reductase inhibitory activity which is also very important because it is the enzyme which is required for the conversion of less harmful testosterone to more harmful dihydrotestosterone in the body, which further cascades a series of events leading to BPH and possibly prostate cancer.
We are also using all of the above mentioned lines.
Are you looking to research the biology of prostate cancer or develop a therapy?
Are you interested in primary or malignant disease? If the latter, for which there is little effective treatment, be aware that androgen independance may be less of an issue, as in many cases, the cells become androgen independant. Most would agree that PC-3 cells, while being easier to grow both in vitro and in vivo only represent the biology of 10-15% of prostate cancer cells. LNCap are better, but in many hands harder to work with in vivo.
LnCaPs are a good model for androgen dependent disease because they express high levels of PSA, that you can control with anit-androgens. LnCaP are however a model of metastatic prostate cancer so you need to make sure that is ok for you.
22Rv1 are another option, but their dependent on androgen is less, they express a mutated AR, and low levels of PSA. they are more representative of primary disease.
LnCaP are not easy to handle and best grown on polylysine coated plates.
I think that if you want to study 'prostate cancer' then focusing on a single cell line is probably not adequate. Different cell lines may faithfully recapitulate certain aspects of the disease, but none will be an accurate model of the disease in general. Cancers are simply too diverse. So, if you want to study any particular aspect, chose a cell line that conforms to this aspect (e.g. elevated activity of a certain enzyme, a certain mutation, ...). If you want to study the disease in general chose a half dozen or even a dozen cell lines that gives a broader representation of the disease and study all of them at once.
I would agree with Michael and Mario. It is quite important to consider what exactly you want to analyze and in order to understand more about prostate cancer one cell line is not enough. Also PC3 cells don't express the AR anymore so they are really AR signaling independent but there is also a PC3-AR cell line available which are PC3 cells stably transfected with a wtAR expression plasmid. Furthermore, LNCaP cells are 'originally' androgen-dependent but there exist high passage LNCaP cells which don't show this dependence anymore. Another important feature of LNCaP cells is that they express a mutant form of the AR (AR T877A).
There are also further cell lines e.g. V-CaP which are androgen independent but express the wtAR, or C4-2 cells which are derived from LNCaP cells.
So, in general there is not THE ONE cell line available to analyze prostate cancer and also to solely look at the 5 alpha reductase also doesn't represent the devo of prostate cancer arpropriately. The best would always be to take different approaches into account as suggested by Mario.
hi there, i work with VCaPs, Du145 and PC3 cells, to cover both androgen sensitive and insensitive areas. i think VCaPs are a good representation of primary prostate cancer (of course they are form a veterbral metastasis), but w.r.t my work, they have similar signalling. they are difficult to work with! what is it you wnat to look at?
Profile the said cell lines on Oncomine.org or Array express or GEO datasets. Specifically look for any changes in expression profile for 5 Alpha reducatse, this will allow you to determine which cell line to use and what to expect. Also try mining a list of genes associated with 5 alpha reductase on genemania and biogrid. The theme of my answer is profiling a data-mine and follow up with invitro work. This will help you save time and get creative with more data. Cheers, RJ.
I think you should work on different cell lines like PC3 DU145, LnCap and BPH.
I have a recent paper on alphavbeta3 sinaling in prostate cancer.
Well Your Question offers big diversity of Answers..........Basic Research takes you to study the Migration and Invasion Property of Androgen Dependent or Independent Cell Line.......For This You can choose LnCap Cell line but this cell line Is very Much Passage Sensitive After certain Passage it wont give you Proper window to analyse your result. Du145 in Some cases gives better expression than LnCap and window too. PC3 you can use but remember in some cases it has been used as Negative control. Please keep in mind some of these and other property for your Specific study.
Please let me know if you have something very specific question and if you are trying regarding Anchorage Independent growth as well.
Best Wishes
Manvendra.
I am happy to read a diversity of approach among varous researchers here. Basically you need to do what Dsr. Abhijit Banerjee and Rajesh Jagirdar suggested, depending on how much money and time you want to spend. LNCaP in general is an excellent cell line for both Androgen-depletion effect studies as well as for 5-alpha-reductase studies. I would do that as the first step. If you need more global picture of inhibition of 5aR, then you need to do what Dr. Jagirdar suggested, but it requires some sophisticated software, may be you could ask him to help!
Bal Lokeshwar, Maimi
I think you first need to specify your goal and depends on your goal, choose the best cell line that matches. If you are interested in Androgen Receptor (AR) signaling pathway, you need cells that are AR positive. LNCaP cells have wt AR and their growth is dependent on androgen level. There are some variants of LNCaP cells that express AR, but their growth is NOT androgen-dependent (e.g. C4-2 or C4-2b). PC-3 and DU145 are prostate cancer cell lines that do have AR, so they are androgen independent as well. I general, when it comes to studies that are done on cell lines, in most of cases you need more than one cell line to show your result in. Good luck....
Hi,
As evoked by others, you have to specify your goal and needs and then look for different approaches. For instance, to see 5 alpha reductase activity I suggest to use protein extract of human or rat testis because to my knowledge the enzymatic activity of the 5 alpha reductase is very faint to be followed in prostatic cell lines. If you want to see anti-cancer effect of somme moleculs you may use different cell lines (androgen-dependent or independent) to see their proliferation, viability and/or apoptotic or anti-apototic protein expression by WB for instance....
Hi! In my opinion you can start the study by using two cell lines: the androgen-dependent LNCaP and the androgen-independent DU-145 or the PC-3. I agree with the suggestions made by others and I recommend to take care in using a certify LNCaP cell line avoiding many cell divisions in culture that could select cells not responsive to androgens. Then, it will be important to extend the investigations to other prostate cancer cell lines as suggested by others. In this regard, 22Rv1 could be a choice in androgen receptor positive cells. Finally, I think that the PZHPV-7 could provide a good model of human non-tumorigenic prostate cells useful as normal control.
It does depend on what you want to do when you choose a cell line. It isn't all that clear from your questions. If you want to compare 5α-reductase in the different cell lines regarding testosterone metabolism (or any other type of comparison between cell lines) it is a good approach to also add a normal cell line (such as PNT2C2) as a control. This allows you to compare what you observe in the two different types of cancer cell lines - androgen dependent and androgen independent, to each other as well as comparing your data to that obtained in a normal cell line.
It realy depend on what you what to see. We worked for a while with androgen receptor ligands and we used PC-3 as AR independent growing cell system the LNCaP as AR dependent growing cell system during the first 30 passages, the VCaP cell line for unmutated AR (detection of ligand binding using PSA level). There are also a series of receptor binding assays (invitrogen). If you want to have a look at the 5-alpha reductase there are transfected HEK cells with the differend reductase types see ( etablierung eukaryontischer Zellinien zur expression der 5alpha reduktase-isoenzyme I und II, W. Reichert; Hybrid inhibitoren der humanen 5 alpha reduktase, M. Schreiber) sorry that it is in german but may be you can find literatur there that help you.
Use LNCaPs, they would be the best invitro model for your experiment. PC3 and DU145 are bit advanced to LNCaPs. Its always better to start with the basic model.
I think most of the audience suggested you to start with the LNCaP cells, but you have to keep end point in your mind that what are you going to do in future. Since, LNCaP has mutatnt AR and PTEN , so in future depends on your plan if your study is just focusing on AR, try to add LAPC4 and VCAP cell lines in your studies because these cell lines has intact AR and PTEN.
You could use the cancer progression model of androgen dependent LNCaP and its androgen independent derivative C81. C81 express AR but is not androgen responsive.
@Klaes Golman. Can you please elaborate the use of NMR spectroscopy for this purpose as you suggested here.
Regards
- Badges
- Science topic