Pharmaceutical composition for preventing or treating liver cancer
a technology of liver cancer and pharmaceutical composition, applied in the direction of drug compositions, capsule delivery, genetic material ingredients, etc., can solve the problems of liver cancer cells that cannot be differentiated, recurrent nodules are difficult to distinguish, and the ehcc generally exhibits minimal dysplasia and lack of clear invasive or destructive growth, so as to prevent the development of liver cancer and inhibit the metastasis and proliferation of liver cancer cells
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example 1
tal Materials and Methods
[0233]1. Cell Culture
[0234]Human liver cancer cell line (SNU-449) and murine Hepa-1c1c7 liver cancer cell line were obtained from Korean Cell Line Bank (Seoul, Korea). All of the cell lines were cultured in EMEM (American Type Culture Collection, Manassas, Va.), RPMI-1640 or DMEM medium (Lonza, Walkersville, Md.) which is supplemented with 10% fetal bovine serum (FBS, Lonza) and 100 units / mL penicillin-streptomycin
[0235](Invitrogen, Carlsbad, Calif.), in a humidified incubator at 37° C. under 5% CO2 condition.
[0236]2. Synthesis and Transfection of siRNA and dsRNA
[0237]The siRNA and dsRNA used in this experiment were synthesized by
[0238]Lemonex (Seoul, Korea). Further, human BANF1, PLOD3 and SF3B4 expression plasmids subcloning gene ORF sequences (BANF1:NM_003860, PLOD3: NM_001084, SF3B4: NM_005850) in pcDNA3.1 + / C-(K)-DYK plasmid, respectively, were purchased from Genscrip™ (Piscataway, N.J., USA). Transfection was performed using Lipofectamine RNAiMAX or Li...
example 2
of Inhibitory Rate of Indicator Gene Expression by siRNA or dsRNA of the Present Invention
[0330]According to the experimental procedures in EXAMPLE 1-1 to 3, indicator genes of siRNA and dsRNA of the present invention (BANF1 variant 1, BANF1 variant 2, PLOD3, and SF3B4) were analyzed, and the results are shown in Tables 7 to 10 below.
[0331]Referring to Tables 7 to 10 below, it can be seen that all the siRNAs and dsRNAs of the present invention could inhibited the expression of the indicator genes at high inhibitory rates.
TABLE 7Verification of validity of siRNA, dsRNA for inhibitionof human BANF1, transcript variant 1, mRNA (Gene Banknumber: NM_003860.3) expressionBase SEQ ID NOExpression inhibitory rate (%)587.73679.64782.3876.21989.61083.421173.181285.441369.571477.31582.921691.381784.111888.361987.832067.722182.292263.232376.242487.72562.572672.922765.582872.91
TABLE 8Verification of validity of siRNA, dsRNA for inhibitionof human BANF1, transcript variant 2, mRNA (Gene Banknumber...
example 3
ation of Excellent RNA Delivery by Porous Silica Particles
[0332]With respect to Hepa-lcic7 and SNU-449 cell lines in EXAMPLE 1, siRNAs, each of which includes a sense RNA having a sequence shown in Table 11 below and an antisense RNA having a complementary sequence thereto, were subjected to in vitro transfection by the methods described in EXAMPLE 1-2 or 1-8, respectively. Then, expression levels of the corresponding markers of the above siRNAs were measured by Western blotting, and the results are shown in FIG. 1.
[0333]Referring to FIG. 1, when the siRNAs shown in Table 11 were transfected, it can be seen that the markers were effectively inhibited. Specifically, when siRNA was loaded on the porous silica particles and then transfected, the expression inhibitory rate was demonstrated to be higher.
TABLE 111Sense RNAName inSEQ ID NOsequenceFIG. 1Target geneSEQ ID NO: 3115′-Banf1Mouse BANF1 geneCCUCAGCGUUUCAAUCUUUUU-3′SEQ ID NO: 3125′-Plod3Mouse PLOD3 geneCGACUGCAGAAUCUCCUCUUU-3′SEQ ...
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