The Effect On Neuroendocfine Differentiation Of Transient Receptor Potential Vanilloid6(TRPV6) In Androgen-independent Progression Of Prostate Cancer

Objective:To investigate the changes of proliferation and invasiveness of prostate cancercells (LNCaP) and the mechanism of prostate cancer androgen-independentprogression by transfecting small interfering RNA (siRNA) and silencing theTransient Receptor Potential Vanilloid6gene of androgen-dependent prostate cancercells (LNCaP).Methods(1) Androgen-dependent prostate cancer cells(LNCaP)and androgen-independentprostate cancer cells(PC-3) were cultured in vitro. Compared the expressionaldifferences of TRPV6between the two kinds of cell lines. After TRPV6gene wassilenced by siRNA, detected the expressions of TRPV6gene in the LNCaP cells.Meanwhile,screened and identificated the steady-transfected LNCaP cells.(2) There set three groups in our experiment after the transformation wassucessfully accomplished:Positive control group(transformation of siRNA-TRPV6),Negative control group (transformation of siRNA-NC)and Blank control group(joined in transfect agents only). Cell proliferation was detected by MTT, while thechanges of invasiveness was detected by using Transwell method. Investigated theexpressional changes of the neuron-specific enolase (NSE) by quantitative PCR andWestern-Blot in the level of gene and protein.(3) Using SPSS19.0to establish a database and analyze the data statistically.The changes of proliferation、invasiveness and the expression of the neuron-specificenolase (NSE) of the LNCaP cells were analysised after the TRPV6gene wassilenced sucessfully.Result:(1) Results of QPCR showed that:TRPV6gene express both in androgen-dependent prostate cancer cells(LNCaP) and androgen-independent prostate cancercells(PC-3), and there`s no significantly difference (P>0.05). (2) Transfection experiments showed that: compared with the Negative controlgroup and Blank control group, the expression of TRPV6siRNA in Positive controlgroup was lower and there was statistically difference(P <0.05). The results of MTTand Transwell methods showed that: compared with the Negative control group andBlank control group, the proliferation and invasion in positive control group wereinhibited (P <0.05), whereas there was no statistically difference between the negativecontrol group and the blank control group (P>0.05).(3) The results of MTT and Transwell methods showed that: compared with theNegative control group and Blank control group, the expression of NSE in Positivecontrol group was lower and there was statistically difference(P <0.05), whereas therewas no statistically difference between the negative control group and the blankcontrol group (P>0.05).Conclusion: Experiment in vitro showed that: silencing the TRPV6gene cansuppress the proliferation and invasiveness of androgen-dependent prostate cancercells(LNCaP),and also have a certain inhibition for prostate cancer androgen-independent progression via neuroendocfine differentiation.