Prostatic cancer related gene and application thereof
A prostate cancer and gene technology, applied in the biological field, can solve problems such as normal cell damage and cancer cell toxicity
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[0079] The preparation method of the small interfering RNA is not particularly limited in the present invention, including but not limited to: chemical synthesis method, in vitro transcription method and the like. It should be understood that those skilled in the art can conveniently prepare or express the small interfering RNA in various ways after knowing the sequence of the siRNA provided by the present invention. For example, in a preferred embodiment of the present invention, the small interfering RNA is chemically synthesized.
[0080] Small interfering RNA can be prepared in the form of double-stranded nucleic acid, which contains a sense strand and an antisense strand, and these two strands form a double strand only under hybridization conditions. A double-stranded RNA complex can be prepared from separate sense and antisense strands. Thus, for example, complementary sense and antisense strands are chemically synthesized and subsequently hybridized by annealing to pro...
Embodiment 1
[0109] Example 1. Cell culture
[0110] Prostate cancer cell lines LNCaP cells, PC-3, DU145, CL-1 and immortalized prostate stromal cells WPMY-1 were purchased from ATCC. LNCaP cells were cultured in DMEM medium containing 5% FBS and 100 U / ml penicillin and streptomycin. Other cell lines were cultured in RPMI 1640 medium containing 10% FBS and 100U / ml penicillin and streptomycin. All cells were kept at 37°C with 5% CO 2 cultured in an incubator.
Embodiment 2
[0111] Example 2.MPSS detects the expression of NDRG3 in human tissue
[0112] Extract various human tissues (see figure 1 ) total RNA, mRNA was synthesized into cDNA double strands with biotin-labeled oligonucleotide primer (oligo-dT); Digested cDNA fragments were purified; synthesis contained 1.67 × 10 7 A tag vector for a 32bp oligonucleotide fragment; clone the purified cDNA fragment into the tag vector, and amplify the insert by PCR primers on the tag vector. Enzyme digestion and digestion of the PCR product to generate a product containing a cDNA fragment connected to a 32bp tag; the cDNA fragment is connected to the microsphere through tag and anti-tag hybrid connection, and each microsphere can also carry 10 4 -10 5 identical cDNA copies.
[0113] Further digest the cDNA template bound on the microsphere, and use different oligonucleotide adapters to connect with the cDNA template attached to the microsphere. In addition, the 3' end of each adapter also has a fluor...
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